Beclabuvir_HNMR_22636_MedChemExpress
A-366_SDS_MedChemExpress
Inhibitors, Agonists, Screening LibrariesSafety Data Sheet Revision Date:May-24-2017Print Date:May-24-20171. PRODUCT AND COMPANY IDENTIFICATION1.1 Product identifierProduct name :A-366Catalog No. :HY-12583CAS No. :1527503-11-21.2 Relevant identified uses of the substance or mixture and uses advised againstIdentified uses :Laboratory chemicals, manufacture of substances.1.3 Details of the supplier of the safety data sheetCompany:MedChemExpress USATel:609-228-6898Fax:609-228-5909E-mail:sales@1.4 Emergency telephone numberEmergency Phone #:609-228-68982. HAZARDS IDENTIFICATION2.1 Classification of the substance or mixtureGHS Classification in accordance with 29 CFR 1910 (OSHA HCS)H302 Acute toxicity, Oral (Category 4)H400 Acute aquatic toxicity (Category 1)H410 Chronic aquatic toxicity (Category 1)2.2 GHS Label elements, including precautionary statementsPictogramSignal word WarningHazard statement(s)H302 Harmful if swallowed.H410 Very toxic to aquatic life with long lasting effects.Precautionary statement(s)P264 Wash skin thoroughly after handling.P270 Do not eat, drink or smoke when using this product.P273 Avoid release to the environment.P301 + P312 IF SWALLOWED: Call a POISON CENTER or doctor/ physician if you feel unwell.P330 Rinse mouth.P391 Collect spillage.P501 Dispose of contents/ container to an approved waste disposal plant.2.3 Other hazardsNone.3. COMPOSITION/INFORMATION ON INGREDIENTS3.1 SubstancesSynonyms:NoneFormula:C19H27N3O2Molecular Weight:329.44CAS No. :1527503-11-24. FIRST AID MEASURES4.1 Description of first aid measuresEye contactRemove any contact lenses, locate eye-wash station, and flush eyes immediately with large amounts of water. Separate eyelids with fingers to ensure adequate flushing. Promptly call a physician.Skin contactRinse skin thoroughly with large amounts of water. Remove contaminated clothing and shoes and call a physician.InhalationImmediately relocate self or casualty to fresh air. If breathing is difficult, give cardiopulmonary resuscitation (CPR). Avoid mouth-to-mouth resuscitation.IngestionWash out mouth with water; Do NOT induce vomiting; call a physician.4.2 Most important symptoms and effects, both acute and delayedThe most important known symptoms and effects are described in the labelling (see section 2.2).4.3 Indication of any immediate medical attention and special treatment neededTreat symptomatically.5. FIRE FIGHTING MEASURES5.1 Extinguishing mediaSuitable extinguishing mediaUse water spray, dry chemical, foam, and carbon dioxide fire extinguisher.5.2 Special hazards arising from the substance or mixtureDuring combustion, may emit irritant fumes.5.3 Advice for firefightersWear self-contained breathing apparatus and protective clothing.6. ACCIDENTAL RELEASE MEASURES6.1 Personal precautions, protective equipment and emergency proceduresUse full personal protective equipment. Avoid breathing vapors, mist, dust or gas. Ensure adequate ventilation. Evacuate personnel to safe areas.Refer to protective measures listed in sections 8.6.2 Environmental precautionsTry to prevent further leakage or spillage. Keep the product away from drains or water courses.6.3 Methods and materials for containment and cleaning upAbsorb solutions with finely-powdered liquid-binding material (diatomite, universal binders); Decontaminate surfaces and equipment by scrubbing with alcohol; Dispose of contaminated material according to Section 13.7. HANDLING AND STORAGE7.1 Precautions for safe handlingAvoid inhalation, contact with eyes and skin. Avoid dust and aerosol formation. Use only in areas with appropriate exhaust ventilation.7.2 Conditions for safe storage, including any incompatibilitiesKeep container tightly sealed in cool, well-ventilated area. Keep away from direct sunlight and sources of ignition.Recommended storage temperature:Powder-20°C 3 years4°C 2 yearsIn solvent-80°C 6 months-20°C 1 monthShipping at room temperature if less than 2 weeks.7.3 Specific end use(s)No data available.8. EXPOSURE CONTROLS/PERSONAL PROTECTION8.1 Control parametersComponents with workplace control parametersThis product contains no substances with occupational exposure limit values.8.2 Exposure controlsEngineering controlsEnsure adequate ventilation. Provide accessible safety shower and eye wash station.Personal protective equipmentEye protection Safety goggles with side-shields.Hand protection Protective gloves.Skin and body protection Impervious clothing.Respiratory protection Suitable respirator.Environmental exposure controls Keep the product away from drains, water courses or the soil. Cleanspillages in a safe way as soon as possible.9. PHYSICAL AND CHEMICAL PROPERTIES9.1 Information on basic physical and chemical propertiesAppearance Light yellow to yellow (Solid)Odor No data availableOdor threshold No data availablepH No data availableMelting/freezing point No data availableBoiling point/range No data availableFlash point No data availableEvaporation rate No data availableFlammability (solid, gas)No data availableUpper/lower flammability or explosive limits No data availableVapor pressure No data availableVapor density No data availableRelative density No data availableWater Solubility No data availablePartition coefficient No data availableAuto-ignition temperature No data availableDecomposition temperature No data availableViscosity No data availableExplosive properties No data availableOxidizing properties No data available9.2 Other safety informationNo data available.10. STABILITY AND REACTIVITY10.1 ReactivityNo data available.10.2 Chemical stabilityStable under recommended storage conditions.10.3 Possibility of hazardous reactionsNo data available.10.4 Conditions to avoidNo data available.10.5 Incompatible materialsStrong acids/alkalis, strong oxidising/reducing agents.10.6 Hazardous decomposition productsUnder fire conditions, may decompose and emit toxic fumes.Other decomposition products - no data available.11.TOXICOLOGICAL INFORMATION11.1 Information on toxicological effectsAcute toxicityClassified based on available data. For more details, see section 2Skin corrosion/irritationClassified based on available data. For more details, see section 2Serious eye damage/irritationClassified based on available data. For more details, see section 2Respiratory or skin sensitizationClassified based on available data. For more details, see section 2Germ cell mutagenicityClassified based on available data. For more details, see section 2CarcinogenicityIARC: No component of this product present at a level equal to or greater than 0.1% is identified as probable, possible or confirmed human carcinogen by IARC.ACGIH: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by ACGIH.NTP: No component of this product present at a level equal to or greater than 0.1% is identified as a anticipated or confirmed carcinogen by NTP.OSHA: No component of this product present at a level equal to or greater than 0.1% is identified as a potential or confirmed carcinogen by OSHA.Reproductive toxicityClassified based on available data. For more details, see section 2Specific target organ toxicity - single exposureClassified based on available data. For more details, see section 2Specific target organ toxicity - repeated exposureClassified based on available data. For more details, see section 2Aspiration hazardClassified based on available data. For more details, see section 212. ECOLOGICAL INFORMATION12.1 ToxicityNo data available.12.2 Persistence and degradabilityNo data available.12.3 Bioaccumlative potentialNo data available.12.4 Mobility in soilNo data available.12.5 Results of PBT and vPvB assessmentPBT/vPvB assessment unavailable as chemical safety assessment not required or not conducted.12.6 Other adverse effectsNo data available.13. DISPOSAL CONSIDERATIONS13.1 Waste treatment methodsProductDispose substance in accordance with prevailing country, federal, state and local regulations.Contaminated packagingConduct recycling or disposal in accordance with prevailing country, federal, state and local regulations.14. TRANSPORT INFORMATIONDOT (US)This substance is considered to be non-hazardous for transport.IMDGUN number: 3077Class: 9Packing group: IIIEMS-No: F-A, S-FProper shipping name: ENVIRONMENTALLY HAZARDOUS SUBSTANCE, SOLID, N.O.S.Marine pollutant: Marine pollutant.IATAUN number: 3077Class: 9Packing group: IIIProper shipping name: Environmentally hazardous substance, solid, n.o.s.15. REGULATORY INFORMATIONSARA 302 Components:No chemicals in this material are subject to the reporting requirements of SARA Title III, Section 302.SARA 313 Components:This material does not contain any chemical components with known CAS numbers that exceed the threshold (De Minimis) reporting levels established by SARA Title III, Section 313.SARA 311/312 Hazards:No SARA Hazards.Massachusetts Right To Know Components:No components are subject to the Massachusetts Right to Know Act.Pennsylvania Right To Know Components:No components are subject to the Pennsylvania Right to Know Act.New Jersey Right To Know Components:No components are subject to the New Jersey Right to Know Act.California Prop. 65 Components:This product does not contain any chemicals known to State of California to cause cancer, birth defects, or anyother reproductive harm.16. OTHER INFORMATIONCopyright 2017 MedChemExpress. The above information is correct to the best of our present knowledge but does not purport to be all inclusive and should be used only as a guide. The product is for research use only and for experienced personnel. It must only be handled by suitably qualified experienced scientists in appropriately equipped and authorized facilities. The burden of safe use of this material rests entirely with the user. MedChemExpress disclaims all liability for any damage resulting from handling or from contact with this product.Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852, USA。
线粒体呼吸链复合体Ⅳ 细胞色素 C 氧化酶活性检测试剂盒说明书
线粒体呼吸链复合体Ⅳ/细胞色素C 氧化酶活性检测试剂盒说明书微量法注意：本产品试剂有所变动，请注意并严格按照该说明书操作。
货号：BC0945规格：100T/96S产品组成：使用前请认真核对试剂体积与瓶内体积是否一致，有疑问请及时联系索莱宝工作人员。
试剂名称规格 保存条件 提取液液体75 mL×2瓶 2-8℃保存 试剂一液体33mL×1瓶 2-8℃保存 试剂二粉剂×2瓶 -20℃保存 试剂三粉剂×2支 2-8℃保存溶液的配制：1、 试剂二：试剂放于试剂瓶内玻璃瓶中。
临用前取1支加入13.5mL 试剂一溶解，用不完的试剂-20℃分装保存2周，避免反复冻融；2、 试剂三：试剂置于试剂瓶内EP 管中；临用前取1支加入2mL 试剂一溶解，用不完的试剂-20℃保存2周，避免反复冻融；3、 工作液的配制：临用前取0.5mL 试剂三加入到溶解好的4.5mL 试剂二中混合备用（约25T ），或者按比例现用现配。
产品说明：线粒体复合体Ⅳ又称细胞色素C 氧化酶，也是线粒体呼吸电子传递链主路和支路的共有成分，负责催化还原型细胞色素C 的氧化，并最终把电子传递给氧生成水。
还原型细胞色素C 在550nm 有特征光吸收，线粒体复合体Ⅳ催化还原型细胞色素C 生成氧化型细胞色素C ，因此550nm 光吸收下降速率能够反映线粒体复合体Ⅳ酶活性。
Reduced Cytochrome C （550nm ） Oxidized Cytochrome C注意：实验之前建议选择2-3个预期差异大的样本做预实验。
如果样本吸光值不在测量范围内建议稀释或者增加样本量进行检测。
需自备的仪器和用品：可见分光光度计/酶标仪、台式离心机、水浴锅/恒温培养箱、可调式移液器、微量玻璃比色皿/96孔板、研钵/匀浆器/细胞超声破碎仪、冰和蒸馏水。
操作步骤：一、样本处理（可适当调整待测样本量，具体比例可以参考文献）1. 称取约0.1g 组织或收集500万细胞，加入1.0 mL 提取液，用冰浴匀浆器或研钵匀浆。
大动脉炎患者外周血单个核细胞RT-qPCR内参基因的选择
January 2021Vol.41 No.12021 年 1 月 第 41 卷 第 1 期基础医学与临床Basic & Clinical Medicine文章编号：1001-6325 ( 2021 ) 01-0087-06研究论文大动脉炎患者外周血单个核细胞RT-qPCR 内参基因的选择田苡箫，李菁*收稿日期：2019-11-18 修回日期：2020-04-30*通信作者(corresponding author ) ：lijing6515@ （中国医学科学院北京协和医学院北京协和医院风湿免疫科风湿免疫病学教育部重点实验室国家皮肤与免疫疾病临床医学研究中心，北京100032）扌摘要：目的筛选适于在大动脉炎（TAK ）患者和健康人群（HC ）之间比较外周血单个核细胞（PBMC ）中mRNA 表达水平的内参基因。
方法提取PBMC 中的总RNA,应用RT-qPCR ，分别采用geNorm 、NormFinder 、BestKeeper 3种软 件程序，分析 3-glucuronidase ,GAPDH ,ACTB ,SDHA ,HPRT1,RPL13A ,B2M , YWHAZ 和 PKG1 9 个基因的 mRNA 表达稳定性。
以T-bet 、GATA3和RORC 作为目的基因，比较不同稳定性的内参基因对mRNA 相对丰度的影响。
结果geNorm 筛选得到的基因组合为B 2M-SDHA , Nor^nFinder 和BestKeeper 筛选出最稳定的内参基因均为HPRT1 ； 3种方法均显示GAPDH 的稳定性较差。
结论自身免疫病患者在接受免疫抑制药物治疗时,原本稳定表达的基因可能会 上调或下调;在样本量较小时，稳定性更好的内参基因可能更有助于检测组间差异。
关键词：大动脉炎;实时定量聚合酶链式反应;RNA 稳定性；内参基因选择中图分类号:R593.2 文献标志码:AValidation of reference genes for the normalization of the RT-qPCRin peripheral blood mononuclear cells of patients with Takayasu arteritisTIAN Yi-xiao ， LI Jing *(Department of Rheumatology and Immunology , Key Laboratory of Rheumatology and Clinical Immunology , Ministry of Education ,National Clinical Research Center for Dermatologic and Immunologic Diseases ( NCRC-DID ),Peking Union Medical College Hospital , CAMS & PUMC , Beijing 100032, China)Abstract ： Objective To validate proper reference genes for quantitative real-time polymerase chain reaction ( RT-qPCR) used for comparing mRNA expression levels in Takayasu arteritis" (TAK) and healthy controls' ( HC ) pe ripheral blood mononuclear cells ( PBMC ). Methods Total RNA in PBMCs was extracted and used RT-qPCR to determine the profiles of 9 candidate genes , including 0-glucuronidase, GAPDH , ACTB , SDHA , HPRT1, RPL13A , B2M , YWHAZ and PKG1. Then compared their transcription stability by geNorm , NormFinder , and Best Keeper. Afterwards , with T-bet , GATA3 and RORC as the targeted genes , explored the influence of reference genes with different stability on mRNA relative abundance. Results The gene combination of B2M-SDHA was selected bygeNorm , and HPRT1 was the most stable one in analysis results of NormFinder and BestKeeper , while GAPDH was less stable. Conclusions Genes that have been expressed stably may be upregulated or downregulated whenpatients with autoimmune diseases received immunosuppressive drugs. When the sample size is small ， the more sta ble internal reference may facilitate the identification of inter-groups difference.Key words : Takayasu arteritis ； real-time polymerase chain reaction ； RNA stability ； selection of reference gene88基础医学与临床Basic&Clinical Medicine2021.41(1)反转录实时荧光定量聚合酶链式反应（reverse quantitative real-time polymerase chain reaction,RT-qPCR）是目前分析基因表达水平的黄金标准，却经常表现出重复性欠佳的问题，选取合适的内参基因有助于改善这一情况［1］。
美国贝克曼库尔特流式细胞分析仪
美国贝克曼库尔特流式细胞分析仪（Beckman coulter cell）产品型号：Cell Lab Quanta SC当前价格：0.00元产品数量：0新旧程度：全新有效期至：0000-00-00所在地：产品简介：仪器简介：T细胞亚群检测的CD45/CD4/CD8/CD3、CD45/CD56/CD19/CD3；阵发性血红蛋白尿（PNH）检测的CD55、CD59；血小板无力症（GT）检测的CD41、CD61等等详细信息仪器简介：T细胞亚群检测的CD45/CD4/CD8/CD3、CD45/CD56/CD19/CD3；阵发性血红蛋白尿（PNH）检测的CD55、CD59；血小板无力症（GT）检测的CD41、CD61等等。
但对于白血病/淋巴瘤免疫分型，国际上迄今为止也没有统一的抗体组合。
在2000年国际细胞分析学会（ISAC）大会上，临床血细胞计数协会组织了一次国际专家会议，以期对检测血液淋巴系统肿瘤所需最少、最有效的单抗数达成共识。
75%与会者一致认为，对于慢性淋巴系统增殖性疾病（CLD）有9种单抗：CD5,CD19,κ,λ,CD3,CD20,CD23,CD10,CD45对初诊来说是最基本的。
淋巴瘤和CLD相似，需要至少12-16种单抗。
对于急性白血病（AL），75%的与会者认为大约13-15种单抗是最基本的：CD10,CD19,CD79a,CD13,CD33,CD34,CD45,CD2,MPO，CD7,CD14,CD3,HLA-DR等，对初步鉴别白血病系列是必需的。
其他一些（CD16,CD56,CDw65,TdT,cyCD3）可能对某些病例有用。
几乎所有的投票者都认为，要对急性白血病完善分类所需单抗的恰当数量平均为20-24种。
但这些抗体之间组合也是一大难题，目前也无统一规定（如表二）。
大会多数发言者(11/13)指出，对已确诊病人的监护和分期来说，仅需较少单抗。
抗体的质量控制是实验的关键环节。
抗体的质量包括其特异性、灵敏度、精密度。
BCA法测肺动脉高压大鼠肺动脉平滑肌SR膜蛋白浓度
BCA法测肺动脉高压大鼠肺动脉平滑肌SR膜蛋白浓度作者：叶兆伟，李洵，王海燕，李尽哲，承伟【摘要】目的测定肺动脉高压大鼠肺动脉平滑肌膜蛋白浓度。
方法蔗糖梯度离心法对肺动脉平滑肌膜蛋白进行提取;二喹啉甲酸(Bicinchoninic acid，BCA )法测定膜蛋白浓度;G-6-Pase定量试剂盒测定平滑肌肌浆网(SR)标志酶G-6-Pase活性。
结果经测定MCT组和对照组G-6-Pase活性分别为(19.5±2.41)IU/g和(18.35±2.21)IU/g，明显高于肺动脉平滑肌组织匀浆G-6-Pase活性(1.23±0.14)IU/g和(1.15±0.17)IU/g;测定波长为595 nm，在0.025～0.5 mg/ml时，线性关系良好，r=0.997 6。
结论提取的SR有较高纯度;BCA法可以用于平滑肌SR膜蛋白浓度的测定。
【关键词】野百合碱;肺动脉高压; SR; BCA法目前测定蛋白浓度有很多种方法，其中二喹啉甲酸(Bicinchoninic acid，BCA)法是近来广为应用的蛋白定量方法。
其原理与Lowry法相似，即在碱性环境下蛋白质与Cu2+络合并将Cu2+还原成Cu+。
BCA与Cu+结合形成稳定的蓝紫色复合物，在562 nm处有高的光吸收值并与蛋白浓度成正比，据此可测算蛋白质浓度。
与Lowry法相比，BCA法灵敏度高、操作简单，试剂及其形成的颜色复合物稳定性俱佳，并且受干扰物质影响小。
与Bradford法相比，BCA法的显著优点是受杂质的影响较小，故本文选用BCA法对提取的样品进行蛋白浓度测定。
1 材料与仪器 1.1 动物健康成年Wistar大鼠，雄雌兼用，250～300 g，由辽宁医学院实验动物中心提供。
1.2 试剂野百合碱(MCT)、Tris、马来酸、二硫苏糖醇(DTT)、苯甲基磺酰氟(PMSF)(美国Sigma公司);抑肽酶(aprotinin)、亮肽素(leupeptin)(Amresco Solarbio公司);BCA蛋白测量试剂盒(北京天来生物医药科技有限公司);葡萄糖-6-磷酸酶定量试剂盒(广州天河天一医药保健品公司);其它试剂均为国产分析纯。
WHO International Standard 1st WHO International Standard for Human Papillomavirus (HPV) Type 16 DNA
WHO International Standard1st WHO International Standard for Human Papillomavirus (HPV)Type 16 DNA NIBSC code: 06/202 Instructions for use(Version 2.0, Dated 10/11/2010)1. INTENDED USEThe 1st International Standard for HPV Type 16 (HPV-16) DNA Nucleic Acid Amplification Techniques consists of a freeze-dried preparation of recombinant plasmid containing full-length HPV-16 DNA cloned via its unique BamH1 site (Quint et al., 2006). The standard has been formulated in a background of purified human genomic DNA, lyophilized in 0.5 ml aliquots and stored at -20 °C. The material was calibrated in an international collaborative study involving 19 laboratories (Wilkinson et al., 2008). The International Standard contains material that is proprietory to third parties and should be used for the sole purpose of calibrating in-house or working standards for the amplification and detection of HPV-16 DNA. The International Standard should not be used for any other purpose and should be discarded after use. 2. CAUTIONThis preparation is not for administration to humans .This material contains DNA derived from C33A cells. As with all materials of biological origin, this preparation should be regarded as potentially hazardous to health. It should be used and discarded according to your own laboratory's safety procedures. Such safety procedures should include the wearing of protective gloves and avoiding the generation of aerosols. Care should be exercised in opening ampoules or vials, to avoid cuts.3. UNITAGEThe 1st International Standard for HPV-16 DNA Nucleic Acid Amplification Techniques has been assigned a unitage of 5 x 106 International Units (IU) per ampoule.Traceability statement:It was proposed at a WHO meeting in January 2008 (WHO Meeting Report, 2008) that the instructions for use of the International Standard for HPV-16 DNA include the calculations and assumptions used in determining the theoretical HPV-16 qenome equivalents (GEq) of the bulk material used in formulating the International Standard, thus demonstrating that 1 IU is equivalent to 1 GEq for HPV-16 DNA . The definitive unitage of the 1st WHO International Standard for HPV-16 DNA therefore remains as IU while the traceability statement would allow users to equate IU with GEq.Assays for DNA concentration of the recombinant HPV-16 plasmid stock preparation were performed in Dr Cosette Wheeler‟s laboratory, University of New Mexico (UNM). DNA concentrations were determined by absorbance at 260 nm as well as spectrofluorometrically using the Picogreen assay (Invitrogen Corporation, USA). A correlation coefficient of 0.95 or higher was obtained between the two DNA measurements. 10 ng HPV-16 plasmid DNA/μl was supplied to NIBSC for formulating the bulk material for subsequent freeze-drying. The UNM laboratory also provided NIBSC with a statement indicating that 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 10 ng HPV-16 plasmid DNA/μl plasmid stock preparation is therefore equivalent to 8.547 x 1011 HPV-16 GEq/ml. NIBSC used this data in formulating the 1st International Standard for HPV Type 16 DNA.Formulation of bulk material for the 1st International Standard for HPV Type 16 DNA (NIBSC code 06/202):At NIBSC, the bulk HPV-16 plasmid DNA material was prepared according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.Therefore,HPV-16 GEq/ml of bulk material = (8.547 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 1.0 x 107 HPV-16 GEq/ml bulk materialThe HPV-16 DNA bulk material was subsequently freeze-dried in 0.5 ml aliquots.Certain assumptions are required for equating IU to GEq for the 1st International Standard for HPV-16 DNA: 1) 1.0 x 1011 GEq/ml for HPV-16 is equal to 1.17 ng/μl. 2) There is no loss in activity of the HPV-16 DNA upon lyophilization. 3) The recombinant HPV-16 plasmid DNA accurately mimics the activity of HPV-16 viral DNA in biological samples.Independent calculation of GEq/ml for recombinant HPV-16 plasmid DNA.NIBSC also independently calculated the genome equivalence of the HPV-16 plasmid stock preparation and bulk preparation in which the molecular weights of the full-length HPV-16 genome and pBR322 DNA were based on sequence content using BioEdit Sequence Alignment Editor v7.0.5.3 (Tom Hall, Isis Pharmaceuticals Inc., USA). The sequences used for determining the molecular weights are GenBank Accession number J01749.1 for pBR322 and the reference sequence for HPV16 (Accession K02718).BioEdit dataDNA molecule: HPV16 Accession K02718 Length = 7904 base pairsMW= 4786756.00 Daltons, double strandedDNA molecule: cloning vector pBR322 Length = 4361 base pairsMW= 2653867.00 Daltons, double strandedFormulaeGEq/ml of the HPV plasmid stock was calculated according to the formula: GEq/ml of the HPV plasmid stock = (DNA concentration of HPV plasmid stock) x (MW of HPV DNA + MW of pBR322)-1 x (Avogadro‟s Number) where Avogadro‟s Number = 6.022x1023 molecules/molGEq/ml of the bulk HPV DNA materials was calculated according to the formula:HPV GEq/ml of bulk material = (HPV GEq/ml of plasmid stock x volume plasmid stock) / volume bulk material.CalculationThe recombinant HPV-16 plasmid stock preparation was supplied to NIBSC at a concentration of 10 ng/μl. Using the MW determinations shown above, the GEq/ml of the HPV-16 plasmid stock is:= (10 x 10-9 g/μl) x (mol/(7440623 g) x (6.022x1023 molecules/mol) = 8.093 x 108 molecules/μl = 8.093 x 1011molecules/ml = 8.093 x 1011 HPV-16 GEq/ml22.23μl of the recombinant HPV-16 plasmid stock was diluted to a final volume of 1900ml, therefore,HPV-16 GEq/ml of bulk material = (8.093 x 1011 HPV-16 GEq/ml plasmid stock) x (0.02223 ml HPV-16 plasmid stock) / 1900 ml HPV-16 bulk material = 0.947 x 107 HPV-16 GEq/ml bulk material4. CONTENTSCountry of origin of biological material: United Kingdom.Each ampoule contains the lyophilized equivalent of 0.5 ml HPV-16 plasmid DNA in 10mM Tris buffer pH7.4 containing 1mM EDTA, 5 mg/ml trehalose and ~1 x 106 human GEq/ml derived from C33a cells.5. STORAGEThe ampoule should be stored at -20 °C or below on receipt.Please note: because of the inherent stability of lyophilized material, NIBSC may ship these materials at ambient temperature.6. DIRECTIONS FOR OPENINGDIN ampoules have an …easy -open‟ coloured stress point, where the narrow ampoule stem joins the wider ampoule body.Tap the ampoule gently to collect the material at the bottom (labeled) end. Ensure that the disposable ampoule safety breaker provided is pushed down on the stem of the ampoule and against the shoulder of the ampoule body. Hold the body of the ampoule in one hand and the disposable ampoule breaker covering the ampoule stem between the thumb and first finger of the other hand. Apply a bending force to open the ampoule at the coloured stress point, primarily using the hand holding the plastic collar.Care should be taken to avoid cuts and projectile glass fragments that might enter the eyes, for example, by the use of suitable gloves and an eye shield. Take care that no material is lost from the ampoule and no glass falls into the ampoule. Within the ampoule is dry nitrogen gas at slightly less than atmospheric pressure. A new disposable ampoule breaker is provided with each DIN ampoule.7. USE OF MATERIALNo attempt should be made to weigh out any portion of the freeze-dried material prior to reconstitution.The 1st International Standard for HPV-16 DNA contains high copy number template. There is a high risk of HPV-16 plasmid DNA contamination via aerosolization upon opening of the glass ampoule. The material must be opened and handled in a separate laboratory environment, away from other pre-amplification components such as reagents, labware and samples.The material is supplied lyophilized and, before use, should be reconstituted in 0.5 ml sterile nuclease-free water. Ensure that the inside surface of the ampoule is wetted with the added water so that any particles of freeze-dried material adhering to the glass are reconstituted. The reconstituted material has a final concentration of 1 X 107 IU/ml. The reconstituted material is suitable for calibration of in-house or working standards for the amplification and detection of HPV-16 DNA.. The material is not suitable for calibrating or assessing extraction, precipitation or centrifugation procedures. The material has NOT been calibrated for human DNA nucleic acid amplification techniques.8. STABILITYReference materials are held at NIBSC within assured, temperature-controlled storage facilities. The 1st International Standard for HPV-16 DNA should be stored at -20 °C or below on receipt.Studies on the stability of reconstituted standard are underway. Users should determine the stability of the reconstituted material according to their own method of preparation, storage and use.NIBSC follows the policy of WHO with respect to its reference materials.9. REFERENCESQuint, W. G. V., Pagliusi, S. R., Lelie, N., de Villiers, E. M., Wheeler, C. M. and the World Health Organization Human Papillomavirus DNA International Collaborative Study Group. (2006). Results of the First WorldHealth Organization International Collaborative Study of Detection of Human Papillomavirus DNA. J. Clin. Microbiol. 44: 571-579.Wilkinson, D.E., Baylis, S.A., Padley, D., Heath, A.B., Ferguson, M., Pagliusi, S.R., et al. Establishment of the 1st World Health Organization international standards for human papillomavirus type 16 DNA and type 18 DNA. Int J Cancer 2010 Jun 15;126(12):2969-83.WHO meeting report, on “Standardization of HPV assays and the role of HPV LabNet in supporting vaccine introduction” Geneva, Switzerland, 23-25 January 2008, in preparation.10. ACKNOWLEDGEMENTS11. FURTHER INFORMATIONFurther information can be obtained as follows; This material: enquiries@ WHO Biological Standards:http://www.who.int/biologicals/en/JCTLM Higher order reference materials: /en/committees/jc/jctlm/ Derivation of International Units:/products/biological_reference_materials/frequently _asked_questions/how_are_international_units.aspx Ordering standards from NIBSC:/products/ordering_information/frequently_asked_q uestions.aspxNIBSC Terms & Conditions:/terms_and_conditions.aspx12. CUSTOMER FEEDBACKCustomers are encouraged to provide feedback on the suitability or use of the material provided or other aspects of our service. Please send any comments to enquiries@13. CITATIONIn all publications, including data sheets, in which this material is referenced, it is important that the preparation's title, its status, the NIBSC code number, and the name and address of NIBSC are cited and cited correctly.15. LIABILITY AND LOSSInformation provided by the Institute is given after the exercise of all reasonable care and skill in its compilation, preparation and issue, but it is provided without liability to the Recipient in its application and use. It is the responsibility of the Recipient to determine the appropriateness of the standards or reference materials supplied by the Institute to the Recip ient (“the Goods”) for the proposed application and ensure that it has the necessary technical skills to determine that they are appropriate. Results obtained from the Goods are likely to be dependant on conditions of use by the Recipient and the variability of materials beyond the control of the Institute.All warranties are excluded to the fullest extent permitted by law, including without limitation that the Goods are free from infectious agents or that the supply of Goods will not infringe any rights of any third party.The Institute shall not be liable to the Recipient for any economic loss whether direct or indirect, which arise in connection with this agreement.The total liability of the Institute in connection with this agreement, whether for negligence or breach of contract or otherwise, shall in no event exceed 120% of any price paid or payable by the Recipient for the supply of the Goods.If any of the Goods supplied by the Institute should prove not to meet their specification when stored and used correctly (and provided that the Recipient has returned the Goods to the Institute together with written notification of such alleged defect within seven days of the time when the Recipient discovers or ought to have discovered the defect), the Institute shall either replace the Goods or, at its sole option, refund the handling charge provided that performance of either one of the above options shall constitute an entire discharge of the Institute‟s liability under this Condition.。
采用重叠PCR构建高灵敏度酵母细胞传感器评估遗传毒性化合物_NormalPdf
1. 1 质粒与菌株 酵母重组质粒 pRNR2-yEGFP 由扬州大学医学
院预防医学系李湘鸣教授赠送；大肠埃希菌 DH5α 感受态细胞（上海生工生物工程有限公司）；酿酒 酵母 BY4741、pESC-Leu 质粒、pESC-His 质粒（上海 柯雷生物科技有限公司）。 1. 2 试 剂
SD/-Ura 、SD/-Leu 、SD/-His 、SD/-His-Leu 、SD/ -Ura-Leu、SD/-Ura-His、SD/-Ura-His-Leu 培养基（上 海艾礼生物科技有限公司）；PrimeSTAR Max DNA 聚合酶（日本 TaKaRa 生物股份有限公司）；PCR 产 物纯化试剂盒、无毒核酸染料、TE 缓冲液、TAE 缓
学报
236
Journal of China Pharmaceutical University 2021，52（2）：236 - 244
采用重叠 PCR 构建高灵敏度酵母细胞传感器 评估遗传毒性化合物
何 颖 1，2，夏星雅 1，2，魏嘉利 1，郑 枫 1，2*
（1中国药科大学药物分析学教研室; 2教育部药品安全与预警重点实验室，南京 210009）
通过采用细胞壁合成与药物转运突变体改善 酵母细胞渗透性，已经实现酵母细胞传感器的检 测灵敏度与特异性的提高，敲除细胞壁合成基因 erg6、cwp1 和 cwp2 与膜转运蛋白基因 pdr5、snq2 和 yor1 是提高酵母细胞传感器检测遗传毒性灵敏度 的有效方法 。 ［18-19］ 酵母细胞具有较高的同源重组 频率和较短的同源片段长度需求，因此采用 40 bp 的同源臂的一步 PCR 产物转化能够进行目标基因 的 敲 除［20］。 为 了 提 高 酵 母 细 胞 基 因 敲 除 的 效 率 ， 需要设计更长的同源臂，而一步 PCR 产物转化法 难以满足，通常需要采取酶切、连接的方法构建基 因 敲 除 组 件［19］。 本 研 究 采 用 重 叠 PCR（overlap PCR）构建 pdr5 与 snq2 基因敲除组件，设计的同源 臂长度为 600~700 bp，相较于传统方法具有成本 更低、操作简便的优点。构建了 pRNR2 调控的基 因突变型酵母细胞传感器对遗传毒性化合物进行 定量评估，并比较了不同基因突变对酵母细胞传 感器检测准确度与灵敏度的影响。
15523666_迷迭香酸对哮喘小鼠氧化性肺损伤的保护作用
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摘要为评价迷迭香酸对哮 喘 小 鼠 模 型 氧 化 性 肺 损 伤 的 保 护 作 用"本 研 究 用 卵 清 蛋 白 %,(;'致 敏$激 发 雌 性 &;B&&S小鼠建立哮喘模型"并用 ,(; 和 Z"," 联合激发小鼠作为氧化肺损伤阳性对照模型#在最后一次滴鼻激 发 "$O 后 "取 支 气 管 肺 泡 灌 洗 液 %&;B['进 行 细 胞 计 数 并 测 定 活 性 氧 %9,D'$超 氧 化 物 歧 化 酶 %D,''和 谷 胱 甘 肽 过 氧化物酶%1DZ?CQ'水平"取左侧肺脏固定做 Z= 染色#结果显示"迷迭香酸可明显减少 &;B[ 中 细 胞 总 数 和 嗜 酸 性粒细胞数目"显著抑制肺组织和 &;B[ 中 9,D的产生"升高 D,' 和 1DZ?CQ水 平"改 善 肺 组 织 病 理 变 化# 本 试 验 结 果 表 明 "迷 迭 香 酸 对 氧 化 肺 损 伤 起 到 明 显 的 保 护 作 用 # 关 键 词 迷 迭 香 酸 *哮 喘 *氧 化 性 肺 损 伤 *抗 氧 化 中 图 分 类 号 9"0434 文 献 标 识 码 ; 文 章 编 号 !+6!?6":+%"8!6'!"?:+48?8+
Beckman Coulter 产品说明书
Product No. C-1432Certified BlankThe Certified Blank reagent contains unlabeled synthetic microspheres of uniform size. These serve as a qualitative control to determine the "noise" level of individual flow cytometers. Certification verifies the detection of the blank beads in fluorescence channels below that of unstained human leukocytes. The product is offered in buffered saline with stabilizer and 0.1% sodium azide (see MSDS)* as preservative. DescriptionStandardization and reproducibility of instrument performance is critical in flow cytometry.1-8 There are a variety of means for monitoring instrument parameters. It is necessary to define fluorescein threshold in order to ensure that a given flow cytometer has the necessary sensitivity for immunophenotyping of cell surface antigens expressed at low density. Instruments should be maintained at a sensitivity level greater than that detected for autofluorescent cells. Fluorescence threshold is defined as the level above which a fluorescence signal can be measured by an instrument. Non-fluorescent particles can be used to make such determinations.ProcedureMaterials required but not provided: Appropriately sized test tubes.Unstained human peripheral blood with eryth-rocytes lysed using a commercial lysing re-agent.Instrumentation required but not provided:Flow cytometer.Optically align flow cytometer per manufacturer’s specifications.Other optional instrument performance QC proce-dures:If FITC/PE two color analysis is routine practice in the laboratory, adjustfluorescence PMT (Photomultiplier tube)and color compensation usingestablished reagent(s)/protocol or FlowCytometry Compensation Kit (SigmaStock No. COMP-1).Analyze Microbead Standards (Sigma Product No. M-0162). Record the PMTvoltages and mean channel fluorescenceon an appropriate log sheet. The resultsshould be consistent with establishedtarget conditions and target channels.The use of Microbead Standards permitsthe direct comparison of fluorescenceintensity data over time.Vigorously shake Certified Blank to achieve a uniform suspension.Add 0.5 ml (approximately 10-15 drops or 200,000 microspheres) to an appropriately sized test tube.Collect data gated on singlets of the Certified Blank for 10,000 events.Record the peak (median) channel for each of the fluorescence parameters. Mean channel may be alternatively used, however, note that mean and median channels are not inter-changeable.Run the lysed, unstained whole blood sample on the flow cytometer, set a gate for lymphocytes and collect 10,000 events.1.Record the mean or median channel at eachPMT.pare the mean or median channels of theunstained human lymphocytes to that of the Certified Blank. If the mean channels of the unstained lymphocytes is above the values for the Certified Blank microbeads, the instrument has sufficient sensitivity for immunophenotyping assay.ProductInformationResultsThe Certified Blank should appear below (to the left of) unstained leukocytes. Each lot is accompanied by a lot-specific certificate verifying the expected mean channel values for a properly calibrated instrument. The Certified Blank serves as a qualitative control for monitoring flow cytometer sensitivity and the level of background fluorescent signals.Figure 1Fluorescence of the Certified Blank in the FITC PMT.Figure 2Autofluorescence of unstained lymphocytes in the FITC PMT.Figure 3Fluorescence of the Certified Blank in the PE PMT.Figure 4Autofluorescence of unstained lymphocytes in the PE PMT.LimitationsProper storage (2-8?C) and handling are essen-tial.This reagent is extremely sensitive to slight changes in pH. The Certified Blank microspheres are maintained at physiological pH (7.2). It is, therefore, critical that pH be carefully controlled in performing quantitative and qualitative flow cyto-metry analyses. Vigorously mixing microspheres prior to use is essential in obtaining a uniform suspension. Establishment of consistent, reproducible mean channel performance data for an appropriate reference standard, i.e., Microbead Standards, is crucial to proper interpretation of results obtained with Certified Blank.StorageStore at 2-8E C. Do Not Freeze.*Due to the sodium azide content a material safety data sheet (MSDS) for this product has been sent to the attention of the safety officer of your institution. Consult the MSDS for information regarding hazards and safe handling practices.References1.Proposed Guidelines: Clinical Applicationsof Flow Cytometry. Quality Assurance andImmunophenotyping of Peripheral BloodLymphocytes. National Committee forClinical Laboratory Standards. DocumentH42-P, 9, 13 (1989).1.Shapiro, H., Practical Flow Cytometry, 2ndEd., Alan R. Liss, Inc., New York, 267-270(1988).2.Keren, D., Flow Cytometry in Clinical Diag-nosis, ASCP Press, Chicago, 27-28 (1989).3.Horan, P., and M. Loken, "A Practical Guideto the Use of Flow Systems", In: Flow Cyto-metry Instrumentation and Data Analysis, M.Van Dilla, et al., (Eds.), Academic Press,NY, 260-280 (1985).4.Horan, P., and J. Kappler, J. Immunol.Meth., 18, 309 (1977).5.McCoy, J., et al., Am. J. Clin. Pathol., 93,Suppl. 1:S27-S37 (1990).6.Giorgi, J.V., et al., Clin. Immunol. Immunop-athol., 55, 173 (1990).7.Parker, J.W., et al., Clin. Immunol. Immuno-pathol., 55, 187 (1990).Sigma warrants that its products conform to the information contained in this and other Sigma publications. Purchaser must determine the suitability of the product for its particular use. See reverse side of invoice or packing slip for additional terms and conditions of the sale. Issued 08/95.。