人白介素18(IL-18)ELISA分析检测试剂盒使用说明书

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白细胞介素18

白细胞介素18白细胞介素18（Interleukin-18）是一种细胞因子，属于白细胞介素家族的一员。

它被认为在免疫应答中起着重要角色，并且与很多疾病的发展和进展有关。

本文将详细介绍白细胞介素18的功能、调控机制以及其在某些疾病中的作用。

白细胞介素18最初是在1995年被发现的，它由肝脏和肝炎病毒感染的肝细胞产生，并被发现能够刺激白细胞产生干扰素-γ（IFN-γ）。

进一步的研究发现，白细胞介素18的产生受到多个因素的调控，包括细胞因子、信号通路和转录因子。

白细胞介素18在免疫应答中的功能是多样的。

首先，它能够刺激NK细胞的活化和增殖，进而促进它们产生INF-γ和其他促炎因子。

其次，白细胞介素18还能够激活巨噬细胞，并增强它们对细菌等外源性病原体的杀伤能力。

此外，白细胞介素18也参与了免疫细胞的极化和调控，对炎症过程的进展具有重要的影响。

白细胞介素18的调控机制非常复杂，它的产生受到多种信号通路的调控。

一些细胞因子，如肿瘤坏死因子（TNF）和白细胞介素1（IL-1）等，能够刺激白细胞介素18的产生。

另外，TLR（Toll-like receptor）信号通路和NLRP3炎症小体也参与了白细胞介素18的调控。

此外，一些转录因子，如NF-κB和AP-1，也能够直接或间接地影响白细胞介素18的产生。

白细胞介素18在多种疾病的发展和进展中发挥着重要作用。

首先，它在免疫性疾病中的作用已经得到了大量的研究。

例如，炎症性肠病（IBD）和风湿性关节炎等疾病中，白细胞介素18的高表达与炎症的严重程度和病情的进展有关。

其次，白细胞介素18还参与了一些肿瘤的发展。

研究发现，白细胞介素18能够促进肿瘤细胞的侵袭和转移，并且与肿瘤的预后密切相关。

此外，白细胞介素18还被发现参与了心血管疾病、神经退行性疾病和肾脏疾病等疾病的发展。

在药物治疗方面，研究人员已经尝试使用白细胞介素18作为治疗免疫性疾病和肿瘤的靶点。

一些抗体和小分子化合物已经被开发出来，以抑制白细胞介素18的活性。

elisa说明书

ELISA KitCatalog # KRC2341 (96 tests)KRC2342 (192 tests)RatIL-18Invitrogen Corporation542 Flynn Road, Camarillo, CA 93012Tel: 800-955-6288E-mail: techsupport@Table of ContentsTable of Contents (3)Contents and Storage (4)Introduction (5)Purpose (5)Principle of the Method (5)Background Information (5)Methods (7)Materials Needed But Not Provided (7)Procedural Notes (7)Preparation of Reagents (8)Assay Procedure (9)Typical Data (10)Performance Characteristics (11)Sensitivity (11)Precision (11)Linearity of Dilution (11)Recovery (12)Specificity (12)Expected Values (12)Stimulation Protocols (12)Limitations of the Procedure (13)Appendix (14)Troubleshooting Guide (14)Technical Support (15)References (16)Citations (16)Contents and Storage Storage Store at 2 to 8°C.ContentsReagents Provided96Test Kit192Test KitRt IL-18 Standard, lyophilized, recombinant baculovirus Rt IL-18.Refer to vial label for quantity and reconstitution volume.2 vials 4 vialsStandard Diluent Buffer. Contains 8 mM sodium azide; 25 ml perbottle.1 bottle2 bottles Incubation Buffer. Contains 8 mM sodium azide; 11 ml per bottle. 1 bottle 1 bottle Rt IL-18 High and Low Control, lyophilized, recombinant baculovirusRt IL-18. Refer to vial label for reconstitution volume and range.2 vials 4 vials Rt IL-18 Antibody-Coated Wells, 96 wells per plate. 1 plate 2 plates Rt IL-18 Biotin Conjugate (Biotin-labeled anti-IL-18). Contains 8 mMsodium azide; 11 ml per bottle.1 bottle2 bottles Streptavidin-Peroxidase (HRP), (100x) concentrate. Contains 3.3 mMthymol; 0.125 ml per vial.1 vial2 vialsStreptavidin-Peroxidase (HRP) Diluent. Contains 3.3 mM thymol;25 ml per bottle.1 bottle 1 bottle Wash Buffer Concentrate (25X); 100 mL per bottle. 1 bottle 1 bottle Stabilized Chromogen, Tetramethylbenzidine (TMB); 25 mL per bottle. 1 bottle 1 bottle Stop Solution; 25 mL per bottle. 1 bottle 1 bottle Plate Covers, adhesive strips. 4 6Disposal Note This kit contains materials with small quantities of sodium azide. Sodium azide reacts with lead and copper plumbing to form explosive metal azides. Upon disposal, flush drains with a large volume of water to prevent azide accumulation. Avoid ingestion and contact with eyes, skin and mucous membranes. In case of contact, rinse affected area with plenty of water. Observe all federal, state and local regulations for disposal.Safety All blood components and biological materials should be handled as potentially hazardous. Follow universal precautions as established by the Centers forDisease Control and Prevention and by the Occupational Safety and HealthAdministration when handling and disposing of infectious agents.IntroductionPurpose The Invitrogen Rat Interleukin-18 (Rt IL-18) ELISA is to be used for the quantitative determination of IL-18 in rat serum, EDTA plasma, buffered solution,or cell culture medium. The assay will recognize both natural and recombinant RtIL-18.For Research Use Only. CAUTION: Not for human or animal therapeutic ordiagnostic use.Principle of the Method The Invitrogen Rt IL-18 kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A polyclonal antibody specific for Rt IL-18 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known Rt IL-18 content, control specimens, and unknowns, are pipetted into these wells.During the first incubation, the Rt IL-18 antigen binds to the immobilized (capture) antibody on one site. After washing, a biotinylated monoclonal antibody specific for Rt IL-18 is added. During the second incubation, this antibody binds to the immobilized Rt IL-18 captured during the first incubation.After removal of excess second antibody, Streptavidin-Peroxidase (enzyme) is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a second incubation and washing to remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of Rt IL-18 present in the original specimen.Background Information IL-18, also known as Interferon-gamma Inducing Factor (IGIF), is a cytokine with Mr=18 kDa (157 amino acid residues) produced by macrophages and monocytes, Kuppfer cells, keratinocytes, intestinal epithelial cells, osteoblasts, mouse diencephalon, and adrenal cortical cells of reserpine-treated rats. IL-18 is synthesized as an inactive precursor molecule with Mr=24 kDa which lacks a signal peptide. The IL-18 precursor is cleaved by IL-1 converting enzyme (ICE, Caspase-1), producing the bioactive, mature form. Only the mature, 18 kDa, form of IL-18 is secreted. Cells that respond to IL-18 include Th1-type cells and NK cells.IL-18 exerts several effects on Th1-like cells. IL-18 stimulates Th1 cell proliferation, Fas ligand expression and IL-2R alpha chain expression.IL-18 also works in combination with IL-12 to induce the production of IFN-γ, GM-CSF, and IL-2 by Th1-type cells. Standard bioassays for mIL-18 measure dose dependent IFN-γ production by IL-18 target cells, such as mouse IL-18 receptor transfected KG-1 cells (human myelomonocyte: ATCC CCL246). Immunomodulatory pathways, which include IL-18 stimulation of IFN-γproduction, are under investigation. IFN-γ production by Th1-type cells and NK cells is important in many immune functions, including defense against viral and parasitic infections, enhancement of NK activity, activation of macrophages, enhancement of B cell function including B cell maturation, proliferation and immunoglobulin secretion, enhancement of MHC class I and class II antigen expression, and inhibition of osteoclast activation.MethodsMaterials Needed But Not Provided •Microtiter plate reader (at or near 450 nm) with software•Calibrated adjustable precision pipettes•Distilled or deionized water•Plate washer: automated or manual (squirt bottle, manifold dispenser, etc.) •Glass or plastic tubes for diluting solutions•Absorbent paper towels•Calibrated beakers and graduated cylindersProcedural Notes 1. When not in use, kit components should be refrigerated. All reagents should bewarmed to room temperature before use.2. Microtiter plates should be allowed to come to room temperature beforeopening the foil bags. Once the desired number of strips has been removed, immediately reseal the bag and store at 2 to 8°C to maintain plate integrity.3. Samples should be collected in pyrogen/endotoxin-free tubes.4. Samples should be frozen if not analyzed shortly after collection. Avoid multiplefreeze-thaw cycles of frozen samples. Thaw completely and mix well prior to analysis.5. When possible, avoid use of badly hemolyzed or lipemic sera. If large amountsof particulate matter are present, centrifuge or filter prior to analysis.6. It is recommended that all standards, controls and samples be run in duplicate.7. When pipetting reagents, maintain a consistent order of addition fromwell-to-well. This ensures equal incubation times for all wells.8. Do not mix or interchange different reagent lots from various kit lots.9. Do not use reagents after the kit expiration date.10. Absorbances should be read immediately, but can be read up to 2 hours afterassay completion. For best results, keep plate covered in the dark.11. In-house controls or kit controls, if provided, should be run with every assay. Ifcontrol values fall outside pre-established ranges, the accuracy of the assay is suspect.12. All residual wash liquid must be drained from the wells by efficient aspiration orby decantation followed by tapping the plate forcefully on absorbent paper.Never insert absorbent paper directly into the wells.13. Because Stabilized Chromogen is light sensitive, avoid prolonged exposure tolight. Avoid contact between chromogen and metal, or color may develop.Directions for Washing •Incomplete washing will adversely affect the test outcome. All washing must be performed with the Wash Buffer Concentrate (25X) provided. •Washing can be performed manually as follows: completely aspirate the liquid from all wells by gently lowering an aspiration tip into the bottom of each well.Take care not to scratch the inside of the well. After aspiration, fill the wells with at least 0.4 ml of diluted Wash Buffer. Let soak for 15 to 30 seconds, then aspirate the liquid. Repeat as directed under Assay Procedure. After the washing procedure, the plate is inverted and tapped dry on absorbent tissue. •Alternatively, the diluted Wash Buffer may be put into a squirt bottle. If a squirt bottle is used, flood the plate with the diluted Wash Buffer, completely filling all wells. After the washing procedure, the plate is inverted and tapped dry on absorbent tissue.•If using an automated washer, follow the washing instructions carefully.Preparation of ReagentsDilution of Standard Note Note: Either glass or plastic tubes may be used for standard dilutions.The Rt IL-18 standard was calibrated against a highly purified recombinant baculovirus protein.1. Reconstitute standard to 5,000 pg/ml with Standard Diluent Buffer. Refer tostandard vial label for instructions. Swirl or mix gently and allow to sit for10 minutes to ensure complete reconstitution. It is recommended thatstandard be used within 1 hour of reconstitution.2. Add 0.1 ml of the reconstituted standard to a tube containing 0.400 mlStandard Diluent bel as 1,000 pg/ml Rt IL-18. Mix.3. Add 0.250 ml of Standard Diluent Buffer to each of 6 tubes labeled 500, 250,125, 62.5, 31.3 and 15.6 pg/ml Rt IL-18.4. Make serial dilutions of the standard as described in the following dilutiondiagram. Mix thoroughly between steps.Remaining reconstituted standard should be discarded. Return the Standard Diluent Buffer to the refrigerator.Preparing SAV-HRP Note: Prepare within 15 minutes of usage. The Streptavidin-HRP (100x concentrate) is in 50% glycerol, which is viscous. To ensure accurate dilution, allow Streptavidin-HRP concentrate to reach room temperature. Gently mix. Pipette Streptavidin-HRP concentrate slowly. Remove excess concentrate solution from pipette tip by gently wiping with clean absorbent paper.1. Dilute 10 µl of this 100x concentrated solution with 1 ml ofStreptavidin-HRP Diluent for each 8-well strip used in the assay. Label asStreptavidin-HRP Working Solution.2. Return the unused Streptavidin-HRP concentrate to the refrigerator.# of 8-Well StripsVolume of Streptavidin-HRPConcentrateVolume of Diluent2 20 µl solution 2 ml4 40 µl solution 4 ml6 60 µl solution 6 ml8 80 µl solution 8 ml10 100 µl solution 10 ml12 120 µl solution 12 ml5,000pg/ml1,000pg/ml500pg/ml250pg/ml125pg/ml62.5pg/ml31.3pg/ml15.6pg/mlpg/ml 100 µlDilution of Wash Buffer 1. Allow the Wash Buffer Concentrate (25X) to reach room temperature and mixto ensure that any precipitated salts have redissolved. Dilute 1 volume of the Wash Buffer Concentrate (25X) with 24 volumes of deionized water (e.g.,50 ml may be diluted up to 1.25 liters, 100 ml may be diluted up to 2.5 liters).Label as Working Wash Buffer.2. Store both the concentrate and the Working Wash Buffer in the refrigerator.The diluted buffer should be used within 14 days.Assay Procedure Be sure to read the Procedural Notes section before carrying out the assay. Allow all reagents to reach room temperature before use. Gently mix all liquid reagents prior to use.Note: A standard curve must be run with each assay.1. Determine the number of 8-well strips needed for the assay. Insert these inthe frame(s) for current use. (Re-bag extra strips and frame. Store these in the refrigerator for future use.)2. Add 100 µl of the Standard Diluent Buffer to the zero standard wells. Well(s)reserved for chromogen blank should be left empty.3. Add 100 µl of standards, samples or controls to the appropriate microtiterwells. (See Preparation of Reagents.)4. Add 50 µl of Incubation Buffer to the zero standard wells and to the wellscontaining standards and serum/plasma samples, or 50 µl of Standard Diluent Buffer to the wells containing cell culture samples and controls.Well(s) reserved for chromogen blank should be left empty. Tap gently on the side of the plate to mix.5. Cover plate with plate cover and incubate for 2hours at roomtemperature.6. Thoroughly aspirate or decant solution from wells and discard the liquid.Wash wells 4 times. See Directions for Washing.7. Pipette 100 µl of biotinylated Rt IL-18Biotin Conjugate solution into eachwell except the chromogen blank(s). Tap gently on the side of the plate to mix.8. Cover plate with plate cover and incubate for 1hour at room temperature.9. Thoroughly aspirate or decant solution from wells and discard the liquid.Wash wells 4 times. See Directions for Washing.10. Add 100 µl Streptavidin-HRP Working Solution to each well except thechromogen blank(s). See Preparation of Reagents.11. Cover plate with the plate cover and incubate for 30minutes at roomtemperature.12. Thoroughly aspirate or decant solution from wells and discard the liquid.Wash wells 4 times. See Directions for Washing.13. Add 100 µl of Stabilized Chromogen to each well. The liquid in the wells willbegin to turn blue.14. Incubate for 30minutes at room temperature and in the dark. Note: Donot cover the plate with aluminum foil or metalized mylar. The incubation time for chromogen substrate is often determined by the microtiter plate reader used. Many plate readers have the capacity to recorda maximum optical density (O.D.) of 2.0. The O.D. values should bemonitored and the substrate reaction stopped before the O.D. of the positive wells exceed the limits of the instrument. The O.D. values at 450 nm can only be read after the Stop Solution has been added to each well. If using a reader that records only to 2.0 O.D., stopping the assay after 20 to25 minutes is suggested.15. Add 100 µl of Stop Solution to each well. Tap side of plate gently to mix. Thesolution in the wells should change from blue to yellow.16. Read the absorbance of each well at 450 nm having blanked the platereader against a chromogen blank composed of 100 µl each of Stabilized Chromogen and Stop Solution. Read the plate within 2 hours after adding the Stop Solution.17. Use a curve fitting software to generate the standard curve. A fourparameter algorithm provides the best standard curve fit.18.Read the concentrations for unknown samples and controls from thestandard curve. (Samples producing signals greater than that of the highest standard should be diluted in Standard Diluent Buffer for serum/plasma samples or corresponding medium for cell culture samples and reanalyzed, multiplying the concentration found by the appropriate dilution factor.)Typical Data (Example) The following data were obtained for the various standards over the range of 0 to 1,000 pg/ml Rt IL-18.StandardRt IL-18 (pg/ml)Optical Density(450 nm)1,000 3.03500 1.94250 1.13125 0.6462.5 0.3831.3 0.2215.6 0.150 0.07Performance CharacteristicsSensitivity The minimum detectable dose of Rt IL-18 is < 4 pg/ml. This was determined by adding two standard deviations to the mean O.D. obtained when the zerostandard was assayed 30 times.Precision 1. Intra-Assay PrecisionSamples of known Rt IL-18 concentration were assayed in replicates of 22 todetermine precision within an assay.Sample 1 Sample 2 Sample 3Mean (pg/ml) 93 387 877SD 3.4 17.3 30.4%CV 3.7 4.5 3.5SD = Standard DeviationCV = Coefficient of Variation2. Inter-Assay PrecisionSamples were assayed 22 times in multiple assays to determine precisionbetween assays.Sample 1Sample 2 Sample 3Mean (pg/ml) 89 386 850SD 6.0 17.1 38.4%CV 6.7 4.4 4.5SD = Standard DeviationCV = Coefficient of VariationLinearity of Dilution Rat serum and cell culture samples were serially diluted in Standard Diluent Buffer or RPMI containing 1% fetal bovine serum,respectively,over the range of the assay. Linear regression analysis of samples versus the expected concentration yielded an average correlation coefficient of 0.99.Serum Cell CultureDilutionMeasured(pg/ml)Expected(pg/ml)%ExpectedMeasured(pg/ml)Expected(pg/ml)%Expectedneat 180 - 194 -1/2 87 90 97 90 97 931/4 43 45 96 44 48.5 911/8 28 22.5 124 25.7 24.3 1051/16 14 11 124 13.5 12.1 111Recovery The recovery of Rt IL-18 added to rat serum averaged 90%. The recovery of Rt IL-18 added to EDTA plasma averaged 95%. The recovery of Rt IL-18 added totissue culture medium containing 1% fetal bovine serum averaged 101%, whilethe recovery of Rt IL-18 added to tissue culture medium containing 10% fetalbovine serum averaged 97%.Specificity Buffered solutions of a panel of substances at 100 ng/ml were assayed with the Invitrogen Rt IL-18 kit. The following substances were tested and found to haveno cross-reactivity: human IL-18, rat IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12p70, IL-13,MIP-2, TNF-α, CINC-2β, VEGF, GM-CSF; mouse IL-18. Both E. Coli andbaculovirus derived Rt IL-18 were detectable with this kit.Expected Values Four pools of rat serum and one pool of rat EDTA plasma were evaluated in this assay. The following concentrations were detected:Sample ConcentrationsPool serum 1 23 pg/mlPool serum 2 51 pg/mlPool serum 3 149 pg/mlPool serum 4 7 pg/mlPool EDTA plasma 6.5 pg/mlStimulation Protocols Cell culture supernatants were evaluated in this assay.Rat Whole Blood (WB) cells were cultured in RPMI for 24, 48 or 72 hours either without stimulus or with a blend of LPS (25 mg/ml) and PHA (5 mg/ml), or with a blend of ionomycin (100 ng/ml) and PMA (100 ng/ml). Results are shown below.IL-18 (pg/ml)Stimulus Cell type24hrs 48hrs72hrsNone WB cells <4 <4 <4LPS + PHA WB cells 8 13 20PMA + ionomycin WB cells 8 22 27Rat splenocytes were cultured at different cellular concentrations in RPMI supplemented with 5% FCS for 24, 48, 72 or 96 hours with a blend of LPS (25 mg/ml) and PHA (5 mg/ml). Results are shown below.IL-18 (pg/ml)CellconcentrationCell type24hrs48hrs72hrs96hrs0.25 x 106 cells/ml Splenocytes17.4 27 36 450.8 x 106 cells/ml Splenocytes49 ND 75 892.5 x 106 cells/ml Splenocytes167 212 238 220Limitations of the Procedure Do not extrapolate the standard curve beyond the top standard point; the dose-response is non-linear in this region and accuracy is difficult to obtain. Dilute all samples above the top standard point with Standard Diluent Buffer; reanalyze these and multiply results by the appropriate dilution factor.The influence of various drugs, aberrant sera (hemolyzed, hyperlipidemic, jaundiced, etc.) and the use of biological fluids in place of serum samples have not been thoroughly investigated. The rate of degradation of native Rt IL-18in various matrices has not been investigated. The immunoassay literature contains frequent references to aberrant signals seen with some sera, attributed to heterophilic antibodies. Though such samples have not been seen to date, the possibility of this occurrence cannot be excluded.Appendix Troubleshooting GuideElevated background Cause: Insufficient washing and/or draining of wells after washing. Solution containing either biotin or SAV-HRP can elevate the background if residual is left in the well.Solution: Wash according to the protocol. Verify the function of automated plate washer. At the end of each washing step, invert plate on absorbent tissue on countertop and allow to completely drain, tapping forcefully if necessary to remove residual fluid.Cause: Contamination of substrate solution with metal ions or oxidizing reagents. Solution: Use distilled/deionized water for dilution of wash buffer and use plastic equipment. DO NOT COVER plate with foil.Cause: Contamination of pipette, dispensing reservoir or substrate solution with SAV-HRP conjugate.Solution: Do not use chromogen that appears blue prior to dispensing onto the plate. Obtain new vial of chromogen.Cause: Incubation time is too long or incubation temperature is too high. Solution: Reduce incubation time and/or temperature.Elevated sample/ standard ODs Cause: Incorrect dilution of standard stock solution; intermediary dilutions not followed correctly.Solution: Follow the protocol instructions regarding the dilution of the standard. Cause: Incorrect dilution of the SAV-HRP conjugate.Solution: Warm solution of SAV-HRP concentrate to room temperature, draw up slowly and wipe tip with kim-wipe to remove excess. Dilute ONLY in SAV diluent provided.Cause: Incubation times extended.Solution: Follow incubation times outlined in protocol.Cause: Incubations carried out at 37°C when RT is dictated.Solution: Perform incubations at RT (= 25 ± 2°C) when instructed in the protocol.Poor standard curve Cause: Improper preparation of standard stock solution.Solution: Dilute lyophilized standard as directed by the vial label only with the standard diluent buffer or in a diluent that most closely matches the matrix of your sample.Cause: Reagents (lyophilized standard, standard diluent buffer, etc.) from different kits, either different cytokine or different lot number, were substituted. Solution: NEVER substitute any components from another kit.Cause: Errors in pipetting the standard or subsequent steps.Solution: Always dispense into wells quickly and in the same order. Do not touch the pipette tip on the individual microwells when dispensing. Use calibrated pipettes and the appropriate tips for that device.Weak/no color developsCause: Reagents not at RT (25 ± 2°C) at start of assay.Solution: Allow ALL reagents to warm to RT prior to commencing assay. Cause: Incorrect storage of components, e.g., not stored at 2 to 8°C.Solution: Store all components exactly as directed in protocol and on labels. Cause: Working SAV-HRP solution made up longer than 15 minutes before use in assay.Solution: Use the diluted SAV-HRP within 15 minutes of dilution.Cause: TMB solution lost activity.Solution 1: The TMB solution should be clear before it is dispensed into the wells of the microtiter plate. An intense aqua blue color indicates that the product is contaminated. Please contact Technical Support if this problem is noted. To avoid contamination, we recommend that the quantity required for an assay be dispensed into a disposable trough for pipetting. Any TMB solution left in the trough should be discarded.Solution 2: Avoid contact of the TMB solution with items containing metal ions. Cause: Attempt to measure analyte in a matrix for which the ELISA assay has not been optimized.Solution: Please contact Technical Support for advice when using nonvalidated sample types.Cause: Wells have been scratched with pipette tip or washing tips.Solution: Use caution when dispensing and aspirating into and out of microwells.Poor PrecisionCause: Errors in pipetting the standards, samples or subsequent steps.Solution: Always dispense into wells quickly and in the same order. Do not touch the pipette tip on the individual microwells when dispensing. Use calibratedpipettes and the appropriate tips for that device. Check for any leaks in the pipette tip.Cause: Repetitive use of tips for several samples or different reagents. Solution: Use fresh tips for each sample or reagent transfer.Cause: Wells have been scratched with pipette tip or washing tips.Solution: Use caution when dispensing and aspirating into and out of microwells.Technical SupportContact Us For more troubleshooting tips, information, or assistance, please call, email, or goonline to /ELISA.USA: Invitrogen Corporation 542 Flynn RoadCamarillo, CA 93012 Tel: 800-955-6288E-mail: techsupport@Europe:Invitrogen LtdInchinnan Business Park 3 Fountain DrivePaisley PA4 9RF, UK Tel: +44 (0) 141 814 6100 Fax: +44 (0) 141 814 6117 E-mail: eurotech@References 1. Dinarello, C.A., et al. (1998) Overview of Interleukin-18: more than an interferon-gamma inducing factor. J. Leukoc. Biol. 63(6):658-664.2. Okamura, H., et al. (1995) Cloning of a new cytokine that induces IFN-gammaproduction by T cells. Nature 378(6552):88-91.3. Ushio, S., et al. (1996) Cloning of the cDNA for human IFN-gamma-inducing factor,expression in Escherichia coli, and studies on the biologic activities of the protein. J.Immunol. 156(11):4274-4279.4. Micallef, M.J., et al. (1996) Interferon-gamma-inducing factor enhances T helper 1cytokine production by stimulated human T cells: synergism with interleukin-12 forinterferon-gamma production. Eur. J. Immunol. 26(7):1647-1651.5. Dao, T., et al. (1996) Interferon-gamma-inducing factor, a novel cytokine, enhancesFas ligand-mediated cytotoxicity of murine T helper 1 cells. Cell. Immunol.173(2):230-235.6. Jordan, J.A., et al. (2001) Role of IL-18 in acute lung inflammation. J. Immunol.167(12):7060-7068.Citations 1. Rana, S., et al. (2005) J. Leukoc. Biol. 77(5):719-28.2. Rosenthal, L.A., et al. (2004) Am. J. Respir. Cell. Mol. Biol. 30: 702-709.For an up-to-date and complete list, visit /ELISA or contact TechnicalSupport.Limited Warranty Invitrogen is committed to providing our customers with high-quality goods and services. Our goal is to ensure that every customer is 100% satisfied with our products and our service. If you should have any questions or concerns about an Invitrogen product or service, please contact our Technical Support Representatives. Invitrogen warrants that all of its products will perform according to the specifications stated on the Certificate of Analysis. The company will replace, free of charge, any product that does not meet those specifications. This warranty limits Invitrogen Corporation’s liability only to the cost of the product. No warranty is granted for products beyond their listed expiration date. No warranty is applicable unless all product components are stored in accordance with instructions. Invitrogen reserves the right to select the method(s) used to analyze a product unless Invitrogen agrees to a specified method in writing prior to acceptance of the order. Invitrogen makes every effort to ensure the accuracy of its publications, but realizes that the occasional typographical or other error is inevitable. Therefore Invitrogen makes no warranty of any kind regarding the contents of any publications or documentation. If you discover an error in any of our publications, please report it to our Technical Support Representatives. Invitrogen assumes no responsibility or liability for any special, incidental, indirect or consequential loss or damage whatsoever. The above limited warranty is sole and exclusive. No other warranty is made, whether expressed or implied, including any warranty of merchantability or fitness for a particular purpose.Licensing Information These products may be covered by one or more Limited Use Label Licenses (see the Invitrogen Catalog or our website, ). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.。

人白介素1β (IL-1β)酶联免疫吸附测定试剂盒使用说明书

2022年修订第一版（本试剂盒仅供体外研究使用，不用于临床诊断!）产品货号：E-EL-H0149c产品规格：96T/48T/24T/96T*5Elabscience 人白介素1β(IL-1β)酶联免疫吸附测定试剂盒使用说明书Human IL-1β(Interleukin 1 Beta) ELISA Kit使用前请仔细阅读说明书。

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Copyright ©2021-2022 Elabscience Biotechnology Co.,Ltd. All Rights Reserved目录用途 (3)基本性能 (3)检测原理 (3)试剂盒组成及保存 (4)试验所需自备物品 (5)样品收集方法 (5)注意事项 (6)■ 试剂盒注意事项 (6)■ 样品注意事项 (6)样本稀释方案 (6)检测前准备工作 (7)操作步骤 (8)结果判断 (10)技术资源 (10)典型数据 (10)性能 (11)■ 精密度 (11)■ 回收率 (11)■ 线性 (11)声明 (12)Intended use (13)Character (13)Test principle (13)Kit components & Storage (14)Other supplies required (15)Sample collection (15)Note (16)■ Note for kit (16)■ Note for sample (16)Dilution Method (17)Reagent preparation (17)Assay procedure (18)Calculation of results (20)Technical resources (20)Typical data (20)Performance (21)■ Precision (21)■ Recovery (21)■ Linearity (21)Declaration (22)用途该试剂盒用于体外定量检测人血清、血浆或其他相关生物液体中IL-1β浓度。

夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书

夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书本试剂仅供研究使用标本：体液夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书试验原理：BFGF试剂盒是固相夹心法酶联免疫吸附实验（ELISA）.已知BFGF浓度的标准品、未知浓度的样品加入微孔酶标板内进行检测。

先将BFGF和生物素标记的抗体同时温育。

人碱性成纤维细胞生长因子ELISA 检测试剂盒洗涤后，加入亲和素标记过的HRP。

再经过温育和洗涤，去除未结合的酶结合物，然后加入底物A、B，和酶结合物同时作用。

产生颜色。

颜色的深浅和样品中BFGF的浓度呈比例关系。

夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书自备材料1．蒸馏水。

2．加样器：5ul、10ul、50ul、100ul、200、500ul、1000ul。

3．振荡器及磁力搅拌器等。

安全性1．避免直接接触终止液和底物A、B。

一旦接触到这些液体，请尽快用水冲洗。

2．实验中不要吃喝、抽烟或使用化妆品。

3．不要用嘴吸取试剂盒里的任何成份。

夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书操作注意事项1．试剂应按标签储存，使用前恢复到室温。

稀稀过后的标准品应丢弃，不可保存。

2．实验中不用的板条应立即放回包装袋中，密封保存，以免变质。

3．不用的其它试剂应包装好或盖好。

不同批号的试剂不要混用。

保质前使用。

4．使用一次性的吸头以免交叉污染，吸取终止液和底物A、B液时，避免使用带金属部分的加样器。

5．使用干净的塑料容器配置洗涤液。

使用前充分混匀试剂盒里的各种成份及样品。

6．洗涤酶标板时应充分拍干，不要将吸水纸直接放入酶标反应孔中吸水。

7．底物A应挥发，避免长时间打开盖子。

底物B对光敏感，避免长时间暴露于光下。

避免用手接触，有毒。

实验完成后应立即读取OD值。

8．加入试剂的顺序应一致，以保证所有反应板孔温育的时间一样。

9．按照中标明的时间、加液的量及顺序进行温育操作。

夏科雷登氏结晶蛋白(CLC)ELISA试剂盒说明书样品收集、处理及保存方法1、血清-----操作过程中避免任何细胞刺激。

人白细胞介素 8（IL-8）定量检测试剂盒（ELISA）说明书

本试剂盒只能用于科学研究，不得用于医学诊断。

人白细胞介素8（IL-8）定量检测试剂盒（ELISA）使用说明书【试剂盒名称】人白细胞介素8（IL-8）定量检测试剂盒（ELISA）【试剂盒用途】定量检测人血清、血浆及相关液体样本中白细胞介素8（IL-8）的含量。

【检测原理】本试剂盒采用双抗体两步夹心酶联免疫吸附法（ELISA）。

将标准品、待测样本加入到预先包被人白细胞介素8（IL-8）单克隆抗体透明酶标包被板中，温育足够时间后，洗涤除去未结合的成分，再加入酶标工作液，温育足够时间后，洗涤除去未结合的成分。

依次加入底物A、B，底物（TMB）在辣根过氧化物酶（HRP）催化下转化为蓝色产物，在酸的作用下变成黄色，颜色的深浅与样品中人白细胞介素8（IL-8）浓度呈正相关，450nm波长下测定OD值，根据标准品和样品的OD值，计算样本中人白细胞介素8（IL-8）含量。

【试剂盒组成】1酶标包被板12孔×8条7显色剂A液6mL2标准品0.3mL×6管8显色剂B液6mL320倍浓缩洗涤液25mL9终止液6mL4样本稀释液6mL10说明书1份5特殊稀释液6mL11封板膜2张6酶标试剂6mL12密封袋1个备注：标准品（1号→6号）浓度依次为：120、60、30、15、7.5、3.75pg/mL.【需要而未提供的试剂和器材】1、37℃恒温箱2、标准规格酶标仪3、精密移液器及一次性吸头4、蒸馏水5、一次性试管6、吸水纸【操作步骤】1、准备：从冰箱取出试剂盒，室温复温平衡30分钟。

2、配液：用蒸馏水将20倍浓缩洗涤液稀释成原倍的洗涤液。

3、加标准品和待测样本：取足够数量的酶标包被板，固定于框架上，分别设置标准品孔、待测样本孔和空白对照孔，记录各孔位置，在标准品孔中加入标准品50μL；待测样本孔中先加入待测样本10μL，再加样本稀释液40μL（即样本稀释5倍）；空白对照孔不加。

4、温育：37℃水浴锅或恒温箱温育30min。

il-18分子量

il-18分子量摘要：1.IL-18 分子的概述2.IL-18 分子量的测定方法3.IL-18 分子量的作用与意义4.IL-18 分子量在医学研究中的应用正文：IL-18 分子的概述IL-18 是一种细胞因子，属于IL-1 家族成员之一，主要参与免疫调节、炎症反应和细胞生长等生理过程。

在免疫应答中，IL-18 能够刺激多种免疫细胞的活化、增殖和分泌细胞因子，从而发挥其抗病毒、抗肿瘤等生物学功能。

IL-18 分子量的测定方法IL-18 分子量的测定方法有多种，常见的有分子量测定、SDS-PAGE 电泳、免疫印迹法等。

分子量测定可以通过测量IL-18 分子在特定条件下的沉降速率来推算其分子量。

SDS-PAGE 电泳则是通过分离IL-18 分子，然后根据其在凝胶中的迁移距离推算其分子量。

免疫印迹法则是通过检测IL-18 分子的抗原性，从而推算其分子量。

IL-18 分子量的作用与意义IL-18 分子量对于其生物学功能具有重要意义。

一方面，IL-18 分子量影响其结构稳定性和活性。

另一方面，IL-18 分子量也会影响其与受体的结合效率，进而影响其生物学效应。

因此，研究IL-18 分子量对于理解其生物学功能和疾病发生机制具有重要意义。

IL-18 分子量在医学研究中的应用IL-18 分子量在医学研究中具有广泛的应用，例如在肿瘤免疫治疗中，研究IL-18 分子量可以为优化抗肿瘤治疗方案提供理论依据。

此外，IL-18 分子量也可以作为评估炎症性疾病、自身免疫性疾病等疾病病情和预后的指标。

总之，IL-18 分子量是一个重要的生物学参数，对于理解IL-18 的生物学功能和疾病发生机制具有重要意义。

白介素18

白细胞介素---18(IL---18)高峰生物技术201007040018IL---18原名IFN--γ诱导因子，是从小鼠及人的肝细胞中克隆得到的一种新的细胞因子，1996年正式命名为IL--18.鼠IL--18前体蛋白由192个氨基酸组成，相对分子质量2.4*104。

N端有一结构特殊的35个氨基酸前导序列，缺乏N--糖基化位点和疏水信号肽样位点。

天然蛋白质由157个氨基酸组成，相对分子质量为1.8*104。

人的IL---18基因编码193个氨基酸前体蛋白，与鼠的IL---18有65%同源性。

SDS--PAGE显示人的IL---18相对分子质量为18.3*103。

人的IL---18主要由单核--巨噬细胞产生，小鼠的IL---18主要由枯否细胞和活化的巨噬细胞产生。

1、IL-18的来源在多种器官、组织和细胞中可检测到IL-18的mRNA，如胸腺、肝脏、脾脏、肾脏、胰腺、枯否细胞和活化的巨噬细胞。

未经处理的小鼠表皮细胞恒定地表达低水平的IL-18mRNA。

体内实验表明，经接触性过敏原刺激后，表皮细胞中IL-18mRNA的表达量在刺激早期就明显上调，在刺激后4～6h 达到峰值。

进一步的实验表明角质形成细胞是表皮中IL-18主要的来源［3］。

成骨细胞基质细胞表达IL-18。

在不支持破骨细胞样多核细胞（OCL）形成的基质细胞系细胞中IL-18的mRNA的表达量较高，在支持OCL形成的基质细胞系的细胞中表达较低［7］。

利血平处理和急性的冷应激都可以使大鼠肾上腺表达IL-18mRNA。

原位杂交显示IL-18的mRNA主要集中在肾上腺皮质，尤其是在产生糖皮质激素的网状带和束状带。

在神经垂体也可检测到IL-18的mRNA，应激对于该部位的IL-18诱生作用并不明显。

这些结果显示IL-18可能是一种神经免疫调节子在免疫系统参与应激过程中起重要的作用［4］。

在胚胎和成体的小肠中测得的IL-18表达量相对于其它器官组织是最高的。

ELISA检测试剂盒使用指南

ELISA检测试剂盒使用指南ELISA（酶联免疫吸附试验）是一种常用的免疫学实验方法，用于检测病原体、抗体或其他分子的存在和浓度。

它具有高灵敏度、高特异性、易操作和较低成本的优势，被广泛应用于生物医学研究、临床诊断和免疫学领域。

本篇文章将介绍ELISA检测试剂盒的使用指南，包括实验准备、试剂的使用步骤和结果解读。

一、实验准备1.阅读检测项目的说明书：在使用试剂盒前，仔细阅读说明书，了解试剂的使用方法、灵敏度和特异性等关键信息。

2.样本准备：根据实验要求，准备样本。

如果需要检测血清中的抗体水平，可以采集血液样本，离心分离血清；如果需要检测细胞培养上清中的分子，将上清收集，并使其清晰，避免细胞或杂质的污染。

3.样本预处理：根据实验需求，对样本进行必要的预处理。

例如，可以通过加热、稀释、酶解等方式来处理样本，以改变样本组分和浓度。

4.准备品质控制样品：制备阳性和阴性控制样品，用于评估试剂盒和实验操作的稳定性和准确性。

5.实验器材准备：准备所需实验器材，如酶标板、移液器、微孔板洗涤仪等。

确保器材的干净和完整，并按照说明书要求对其进行预处理。

二、试剂使用步骤1.试剂准备：根据说明书要求，将试剂从冰箱中取出，并在室温下放置一段时间使其恢复到室温。

确保试剂瓶盖紧闭，并避免暴露在直接阳光下。

2.实验操作：按照说明书的要求，将试剂加入到酶标板中，并根据实验设计进行标准曲线的设置。

标准曲线用于测量未知样品的数量，并计算出浓度。

3.孵育：根据试剂盒的要求，将酶标板放入孵育箱中进行孵育。

孵育温度和时间应根据实验要求进行调整。

4.洗涤：使用洗涤缓冲液对酶标板上的不特异性结合物进行洗涤。

洗涤过程应准确控制洗涤孔板次数和洗涤液的体积。

5.补液：在洗涤完成后，加入辣根过氧化物酶标况稀释液，促进酶标物与特异性结合物的反应。

6.孵育：根据试剂盒的要求，将酶标板放入孵育箱中进行二次孵育。

7.反应停止：根据试剂盒的要求，加入相应的停止液，停止酶反应。

ELISA 检测试剂盒说明书

本试剂盒只能用于科学研究，不得用于医学诊断人（Human Human））17-17-羟皮质类固醇（羟皮质类固醇（羟皮质类固醇（17-OHCS 17-OHCS 17-OHCS））ELISA 检测试剂盒使用说明书检测原理试剂盒采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。

往预先包被17-羟皮质类固醇（17-OHCS）抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并彻底洗涤。

用底物TMB显色，TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。

颜色的深浅和样品中的17-羟皮质类固醇（17-OHCS）呈正相关。

用酶标仪在450nm 波长下测定吸光度（OD 值），计算样品浓度。

样品收集、处理及保存方法1.血清：使用不含热原和内毒素的试管，操作过程中避免任何细胞刺激，收集血液后，3000转离心10分钟将血清和红细胞迅速小心地分离。

2.血浆：EDTA、柠檬酸盐或肝素抗凝。

3000转离心30分钟取上清。

3.细胞上清液：3000转离心10分钟去除颗粒和聚合物。

4.组织匀浆：将组织加入适量生理盐水捣碎。

3000转离心10分钟取上清。

5.保存：如果样本收集后不及时检测，请按一次用量分装，冻存于-20℃，避免反复冻融，在室温下解冻并确保样品均匀地充分解冻。

自备物品1.酶标仪（450nm）2.高精度加样器及枪头：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒温箱操作注意事项1.试剂盒保存在2-8℃，使用前室温平衡20分钟。

从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶完全溶解后再使用。

2.实验中不用的板条应立即放回自封袋中，密封（低温干燥）保存。

3.浓度为0的S0号标准品即可视为阴性对照或者空白；按照说明书操作时样本已经稀释5倍，最终结果乘以5才是样本实际浓度。

4.严格按照说明书中标明的时间、加液量及顺序进行温育操作。

5.所有液体组分使用前充分摇匀。

ELISA_kit使用说明书

微囊藻毒素ELISA检测试剂盒使用说明书Microcystin Plate Kit一、基本原理酶联免疫（ELISA）是免疫酶技术的一种，是将抗原抗体反应的特异性与酶反应的敏感性相结合而建立的一种新技术。

ELISA的技术原理是：将酶分子与抗体（或抗原）结合，形成稳定的酶标抗体（或抗原）结合物，当酶标抗体（或抗原）与固相载体上的相应抗原（或抗体）结合时，即可在底物溶液参与下，产生肉眼可见的颜色反应，颜色的深浅与抗原或抗体的量成比例关系，使用ELISA 检测仪，即酶标仪，测定其吸收值可做出定量分析。

此技术具有特异、敏感、结果判断客观、简便和安全等优点，日益受到重视，不仅在微生物学中应用广，而且也被其他学科领域广为采用。

本试剂盒工作原理如下图所示。

二、已配试剂ELISA试剂盒中配备以下条目：1、96孔已包被好的酶标板2、1只阴性对照即0孔溶液3、标准1：0.1ng/mL的MC-LR标准溶液4、标准2：0.2ng/mL的MC-LR标准溶液5、标准3：0.5ng/mL的MC-LR标准溶液6、标准4：1ng/mL的MC-LR标准溶液7、标准5：2.5ng/mL的MC-LR标准溶液8、1瓶溶液1 (单抗)9、1只溶液2(二抗稀释液)10、一只二抗（小管乘装）11、1瓶溶液3（底物溶液）12、1瓶溶液4 （终止液）13、1袋固体PBS（配置洗涤液用）三、需配器材及试剂除试剂盒内的物品外，还需配备以下器材和试剂：1、酶标仪（若可能，可配有洗板机）2、微量移液器（100 L）（若可能，可配8道或12道微量移液器）3、吸管、橡皮吸头（洗板时用，若有洗板机，无需准备此项）4、纯水（用于配置洗涤液）5、玻璃瓶（盛装洗涤液）6、记时器（准确控制每步操作时间）7、封口膜（防止孔内液体挥发）四、酶标二抗的配置将小管中二抗用溶液2按1:3000稀释，充分溶解混匀，现用现配。

五、洗涤液的配置将固体PBS以纯水配置成1L溶液，加1mL Tween 20（PBS-T, pH7.4-7.6），室温保存。

人(Human)细胞角蛋白18-M30(CK18-M30)-NEWA

上海笃玛生物科技有限公司本试剂盒只能用于科学研究，不得用于医学诊断人（Human）细胞角蛋白18-M30（CK18-M30）ELISA 检测试剂盒使用说明书检测原理试剂盒采用双抗体一步夹心法酶联免疫吸附试验（ELISA）。

往预先包被细胞角蛋白18-M30（CK18-M30）抗体的包被微孔中，依次加入标本、标准品、HRP标记的检测抗体，经过温育并彻底洗涤。

用底物TMB显色，TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。

颜色的深浅和样品中的细胞角蛋白18-M30（CK18-M30）呈正相关。

用酶标仪在450nm 波长下测定吸光度（OD 值），计算样品浓度。

2.血浆：EDTA、柠檬酸盐或肝素抗凝。

3000转离心30分钟取上清。

3.细胞上清液：3000转离心10分钟去除颗粒和聚合物。

4.组织匀浆：将组织加入适量生理盐水捣碎。

3000转离心10分钟取上清。

5.保存：如果样本收集后不及时检测，请按一次用量分装，冻存于-20℃，避免反复冻融，在室温下解冻并确保样品均匀地充分解冻。

从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶完全溶解后再使用。

2.实验中不用的板条应立即放回自封袋中，密封（低温干燥）保存。

3.浓度为0的S0号标准品即可视为阴性对照或者空白；按照说明书操作时样本已经稀释5倍，最终结果乘以5才是样本实际浓度。

4.严格按照说明书中标明的时间、加液量及顺序进行温育操作。

上海笃玛生物科技有限公司5.所有液体组分使用前充分摇匀。

白介素-18的正常值

白介素-18的正常值
白介素-18（IL-18）是一种细胞因子，对免疫反应和炎症过程
起着重要作用。

正常情况下，IL-18的浓度可以受到多种因素的影响，包括年龄、性别、生理状态等。

一般来说，IL-18的正常值在
不同的研究和临床实验中可能会有所不同，但通常在健康成年人中，IL-18的正常浓度范围约为100-300 pg/mL。

需要注意的是，IL-18的正常范围可能因不同的实验室方法和
试剂盒而有所差异，因此在进行临床检测时，应该参考具体实验室
提供的参考范围。

此外，IL-18的正常值也可能因个体差异而有所
变化，因此在进行临床诊断时，应该综合考虑患者的临床症状和其
他相关检测结果。

总之，IL-18的正常值是一个相对范围，在临床实践中需要结
合具体情况进行分析和判断。

如果你需要进行IL-18的检测，建议
咨询专业医生以获取更准确的信息。

人淋巴细胞因子ELISA试剂盒说明书

人淋巴细胞因子ELISA试剂盒说明书
人淋巴细胞因子ELISA定量检测试剂盒等ELISA试剂盒检测类型：双抗体夹心法测抗原、双抗原夹心法测抗体、间接法测抗体、竞争法测抗体、竞争法测抗原、捕获包被法测抗体、ABS-ELISA法。

我司人淋巴细胞因子ELISA定量检测试剂盒等ELISA试剂盒，灵敏性高，高效性，原装进口抗体，吸附均匀、吸附性好，回收利用率高、可靠性强，无效包退包换，公司提供免费代测服务，各种种属ELISA 试剂盒产品齐全，质量可靠。

详情可来电咨询销售人员或咨询我司在线客服。

人淋巴细胞因子ELISA定量检测试剂盒操作步骤：
1、加样：分别设空白孔(空白对照孔不加样品及酶标试剂，其余各步操作相同)、标准孔、待测样品孔。

在酶标包被板上标准品准确加样50µl，待测样品孔中先加样品稀释液40µl，然后再加待测样品10µl （样品最终稀释度为5倍）。

加样将样品加于酶标板孔底部，尽量不触及孔壁，轻轻晃动混匀。

2、温育：用封板膜封板后置 37℃温育30分钟。

3、配液：将30倍浓缩洗涤液用蒸馏水30倍稀释后备用
4、洗涤：小心揭掉封板膜，弃去液体，甩干，每孔加满洗涤液，静置30秒后弃去，如此
重复5次，拍干。

5、加酶：每孔加入酶标试剂50µl，空白孔除外。

6、显色：每孔先加入显色剂A50µl，再加入显色剂B50µl，轻轻震荡混匀，37℃避光显色
10 分钟.
7、终止：每孔加终止液50µl，终止反应(此时蓝色立转黄色)。

8、测定：以空白空调零，450nm波长依序测量各孔的吸光度(OD值)。

测定应在加终止液后15分钟以内进行。

鸡白细胞介素-18(ChIL-18)重组蛋白的生物学活性检测

农业生物技术学报Journal of Agricultural Biotechnology 2007,15(1):5~10*基金项目：山东农业大学博士科研基金项目(No.23201)、“十一五”国家科技支撑计划重大项目(No.20061122)和山东省优秀中青年科学家科研奖励基金(No.2006BS06015)资助。

**通讯作者。

Author for correspondence.教授，博士,主要从事畜禽传染病学和细胞因子的研究。

E-mail:<hkzhao@>.收稿日期：2006-01-11接受日期：2006-03-30·研究论文·鸡白细胞介素-18(ChIL-18)重组蛋白的生物学活性检测*李宏梅1,胡敬东1,马凤龙2,郑玉姝1,刘翠艳1,田兆菊1,赵宏坤1**(1.山东农业大学动物科技学院，泰安271018；2.山东信得药业,青岛266061)摘要：对含有鸡白细胞介素-18()基因的质粒在大肠杆菌()内进行诱导表达，经亲和层析法对表达产物进行纯化，用微量细胞病变抑制法CEF-VSV 系统，检测其对SPF 鸡脾淋巴细胞诱导产生γ-干扰素(IFN-γ)的活性和其对病毒的抑制效果；用3H-TdR 掺入法和MTT 法分别检测其对T 淋巴细胞诱导转化和NK 细胞杀伤活性的作用。

结果表明，经诱导表达出分子量44kD 的目的蛋白(与GST 融合表达)，纯化后得到去除菌体蛋白的高纯度目的蛋白；该蛋白能够诱导脾细胞产生IFN-γ，当浓度为250ng/mL 时诱导IFN-γ的活性最高，可达1×106U/mL ；但该蛋白不能直接抑制水疱性口炎病毒(VSV)在鸡胚成纤维细胞(CEF)上生长；能够刺激淋巴细胞显著增殖，浓度为250ng/mL 时效果最佳；适当浓度的rChIL-18蛋白能促进NK 细胞的杀伤活性，浓度为150ng/mL 时，作用最强。

利用原核表达的重组鸡白细胞介素-18蛋白与天然鸡IL-18蛋白一样具有较广泛的生物学活性。

小儿过敏性紫癜血清白介素18含量的检测分析

小儿过敏性紫癜血清白介素18含量的检测分析张国华【摘要】目的:检测并分析过敏性紫癜患儿白介素18的含量.方法:过敏性紫癜患儿58例,根据临床尿检结果分为单纯性紫癜组30例和紫癜性肾炎组28例,另设健康对照组20例;采用双抗体夹心酶联免疫吸附法,检测患儿白介素18的血清水平,并统计进行分析.结果:单纯性紫癜组及紫癜性肾炎组白介素18的检出值均明显高于健康对照组;紫癜性肾炎组的血清白介素18水平高于单纯性紫癜组,差异均有统计学意义.结论:过敏性紫癜患儿血中有较高的白介素18表达;白介素18的表达与患儿的肾脏损害相关.【期刊名称】《中国民族民间医药》【年(卷),期】2013(022)022【总页数】1页(P63-63)【关键词】过敏性紫癜;白介素18;肾脏损害;检测分析【作者】张国华【作者单位】南方医科大学附属佛山妇幼保健院,广州,佛山,528000【正文语种】中文【中图分类】R725.2过敏性紫癜（HSP）多发于儿童，是以坏死性小血管炎为主要病理改变的全身性疾病。

本文检测了HSP患儿血清中白细胞介素18（IL－18）的表达水平。

旨在比较分析IL－18在小儿HSP病程中的表达情况，并探讨它们与肾脏损害的关联性，现报告如下。

1.1 材料选取2009年至2011年佛山市多家医院儿科确诊为HSP的住院患儿58例为研究对象，根据尿检结果分为单纯性紫癜组30例和紫癜性肾炎组28例。

58例患儿中，男25例，女33例；年龄：单纯性紫癜组8.37±2.26岁，紫癜性肾炎组7.89±3.38岁；另设健康对照组20例。

1.2 具体分组及入选标准为1.2.1 健康对照组20例，为本院常规性体检小儿，均无肾脏、风湿及哮喘疾病史。

1.2.2 单纯性紫癜组30例，包含皮肤型、腹型、关节型以及混合型。

诊断标准按《实用儿科学》第7版所列1990年美国风湿病协会制定标准［1］。

1.2.3 紫癜性肾炎组28例，即过敏性紫癜病程中伴有血尿和（或）蛋白尿。

人细胞角蛋白

人细胞角蛋白18-M30（CK18-M30）酶联免疫分析试剂盒使用说明书本试剂盒仅供体外研究使用!预期应用ELISA法定量测定人血清、血浆或其它相关生物液体中CK18-M30含量。

实验原理用纯化的抗体包被微孔板，制成固相载体，往包被抗CK18-M30抗体的微孔中依次加入标本或标准品、生物素化的抗CK18-M30抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。

TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。

颜色的深浅和样品中的CK18-M30呈正相关。

用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。

试剂盒组成及试剂配制1.酶联板：一块（96孔）2.标准品（冻干品）：2瓶，每瓶临用前以样品稀释液稀释至1ml，盖好后静置10分钟以上，然后反复颠倒/搓动以助溶解，其浓度为10ng/ml，做系列倍比稀释（注：不要直接在板中进行倍比稀释）后，分别稀释成10ng/ml，5ng/ml，2.5ng/ml，1.25ng/ml，0.625ng/ml，0.312ng/ml，0.156ng/ml，样品稀释液直接作为标准浓度0ng/ml，临用前15分钟内配制。

如配制5ng/ml标准品：取0.5ml（不要少于0.5ml）10ng/ml 的上述标准品加入含有0.5ml样品稀释液的Eppendorf管中，混匀即可，其余浓度以此类推。

3.样品稀释液：1×20ml。

4.检测稀释液A：1×10ml。

5.检测稀释液B：1×10ml。

6.检测溶液A：1×120μl（1:100）临用前以检测稀释液A1:100稀释，稀释前根据预先计算好的每次实验所需的总量配制（100μl/孔），实际配制时应多配制0.1-0.2ml。

如10μl检测溶液A加990μl检测稀释液A的比例配制，轻轻混匀，在使用前一小时内配制。

7.检测溶液B：1×120μl/瓶（1:100）临用前以检测稀释液B1:100稀释。

病毒性心肌炎患儿血清白介素-18(IL-18)含量变化及临床意义

病毒性心肌炎患儿血清白介素-18(IL-18)含量变化及临床意
义
施耀岳;许英
【期刊名称】《齐齐哈尔医学院学报》
【年(卷),期】2008(29)19
【摘要】目的探讨病毒性心肌炎患儿血清白介素-18(IL-18)的含量变化及其临床意义.方法采用酶联免疫吸附试验ELISA法检测45例病毒性心肌炎患儿及40例正常对照儿童血清中IL-18的表达水平.结果观察组中非死亡组、死亡组患儿血清IL-18含量明显高于对照组患儿(P<0.05),有显著性差异;非死亡组治疗前后血清IL-18含量有显著性差异(P<0.05).结论 IL-18在病毒性心肌炎的发生发展和治疗中具有重要意义.
【总页数】1页(P2320)
【作者】施耀岳;许英
【作者单位】江苏省海门市人民医院,226100;江苏省海门市人民医院,226100【正文语种】中文
【中图分类】R9
【相关文献】
1.乙型脑炎病毒感染患儿血清中IL-12和IL-18的含量变化 [J], 黄友敏;刘松华
2.病毒性心肌炎患儿血清IL-10,IL-18水平的变化及临床意义 [J], 赵洪晶
3.病毒性心肌炎患儿血清白介素23和白介素17水平的变化及其临床意义 [J], 蔡
泽民;杨云华
4.乙脑患儿血清和脑脊液中IL-18含量变化及其临床意义 [J], 刘培华;余光开
5.心肌炎患儿血清中细胞因子IL-18、IFN-γ含量变化的临床意义 [J], 杨楠;张宏艳;陈朝晖;张炜
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