实验室用液体配制

抗体稀释液

牛血清白蛋白1g

NaN3 0.08g

30% Triton 1ml（PBS）

定容PBS（1x）100ml

驴血清封闭液

血清5-10ml

NaN3 0.05g

30% Triton 0.5ml（PBS）

定容PBS（1x）100m

抗原稀释液

A 18ml+

B 82 ml5

双蒸水1000ml ,PH=6

封片液

Na2CO3 0.689g

NaHCO3 1.554g

溶于50ml 双蒸水

50ml 甘油加热混合

抗原修复液

A:柠檬酸2.1g----100ml ddH2O

B:二水柠檬酸三钠14.7g-----500ml

A 18ml+

B 82ml+900mlddH2O--------1L(PH=6.0)

Neurobasal Media

glutamine, 1%penicillin/streptomycin, 10 ng/mL platelet-derived

growth factor, 10 ng/mL FGF, and 2% B27

BRDU

BrdU is a brominated analog of thymidine.

STABILITY / STORAGE AS SUPPLIED:

BrdU should be stored at -0°C and desiccated. If properly stored, it will have a shelf-life of two years.

SOLUBILITY / SOLUTION STABILITY:

Sigma routinely tests the solubility at 50 mg/mL in 1 M ammonium hydroxide yielding a clear colorless solution. It can be dissolved in water at a concentration of up to 10 mg/mL. It is also soluble in DMF, DMSO and water (with heat) at 50-100 mg/mL.

Solutions at neutral pH should be stable for at least 6 months when stored frozen at -20°C. APPLICATIONS:

BrdU is selectively incorporated into cell DNA at the S phase of cell cycle. The use of BrdU as a thymidine analog has made possible the identification of DNA synthesis in suspensions of cells, cell smears and tissue sections. The incorporation of BrdU into DNA

in place of thymidine is discussed by A.L. Givan.BrdU at 0.16 to 500 μg/mL of cell culture media produced inhibition of growth of KD cells (rabbit kidney cells). Effective inhibition at concentrations greater than 1.0 μg/mL was observed.6 It is i ncorporated, in vivo, by injecting 10-100 mg/kg at 10 mg/mL in saline intraperitoneally.7 It is also incorporated into bone marrow cells in culture at a final concentration of 10 μM at 37C for one hour. For incorporation to occur, the BrdU must be phosphorylated in the cell by thymidine kinase. FITC-conjugated second antibodies can be used with antibodies specific for BrdU (Sigma product B2531) which will make "new" DNA fluoresce green; denatured DNA can be stained with propidium iodide and will fluoresce red.

PeproTech细胞因子和重组蛋白溶解必读

来源：美国PeproTech(派普泰克)公司中国代表处

大家知道，PeproTech的所有细胞因子和蛋白均为冻干粉，这使得运输非常便捷，只要常温即可。而且，细胞因子和蛋白冻干粉非常稳定，在-20o C或-80o C条件下可保存数年。冻干粉在使用前需进行溶解，然后以液体形式加到培养体系或注射入动物体内。溶解步骤非常关键，因溶解不好会导致细胞因子或蛋白的失活，这也是很多用户在实际使用中经常遇到的问题。

应该如何进行正确的溶解呢？

下面我们以Recombinant Human IL-4 (重组人IL-4，产品编号：200-04)的说明书为例，对细胞因子或蛋白的溶解方法进行详细的阐述。

拿到重组人IL-4的说明书后，您会发现有一段关于Reconstitution(重悬)的叙述，这段内容含有溶解相关的所有信息。

1. Centrifuge the vial prior to opening

第 1 步：开盖前离心试剂管

PeproTech的细胞因子或蛋白冻干粉装盛在塑料管中，为无菌包装。冻干粉在运输过程中可能会因颠簸而漂散并粘贴于管壁或管盖上，所以在打开塑料瓶盖前，需将冻干粉通过离心收集到管底，以便用很小体积的液体即可将冻干粉完全溶解。

有很多用户会问一个问题，即应该用多少转速、多长时间离心试剂管，才能达到良好的收集效果？

答：有些小型高速离心机(多为进口品牌)的面板上有一个Spin键，按了此键后，离心机会自动快速上升到其最大速度(10000rpm或12000rpm)，上升到最高点后速度即刻下降，直至停止旋转，整个过程大约30s。这个Spin键足以很好的将细胞因子或蛋白收集到管底。

但有些实验室没有这样的高速离心机，只有最高转速为4000-4500rpm的离心机。这种情况下，需3000-3500rpm离心5min，也能达到类似的效果。

2. Reconstitute in water to a concentration of 0.1-1.0 mg/ml. Do not vortex.