BrdU Cell Proliferation Assay Kit

产品编号：CST-6813S 品牌：CST 原厂货号：6813S
产品分类：生化和细胞分析 > 细胞分析试剂盒 > 细胞增殖及周期检测试剂盒
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包装：

运保温度:

-20° C

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其它组分：

STOP Solution 【子货号：#7002，包装：50 ml，，运保温度：-20° C】

TMB Substrate 【子货号：#7004，包装：50 ml，，运保温度：-20° C】

ELISA Wash Buffer (20X)【包装：50 ml，，运保温度：-20° C】

Anti-mouse IgG, HRP-linked Antibody【包装：0.5 ml，，运保温度：-20° C】

BrdU【包装：0.15 ml，，运保温度：-20° C】

HRP-linked Antibody Diluent【包装：50 ml，，运保温度：-20° C】

Detection Antibody Diluent【包装：50 ml，，运保温度：-20° C】

BrdU Detection Antibody【包装：0.5 ml，，运保温度：-20° C】

Fixing/denaturing Solution【包装：25 ml，，运保温度：-20° C】

描述：

BrdU Cell Proliferation Assay kit能够检测细胞增殖过程中掺入到细胞DNA中的BrdU。BrDU标记的DNA变性后能够为试剂盒中的BrdU Mouse mAb检测到。BrdU Mouse mAb不与内源性DNA发生交叉反应。依细胞种类和孵育时间不同，每孔0.2-2x104个细胞即足够大部分实验。为了得到最佳实验结果，我们推荐进行细胞数滴度实验（图1）。BrdU Cell Proliferation Assay kit能够检测细胞增殖过程中掺入到细胞DNA中的BrdU，使用的抗体为抗BrDU抗体。当细胞培养于含BrDU的标记培养基中，该胸腺嘧啶类似物可代替胸腺嘧啶掺入到增殖细胞的新合成DNA中。吸除标记培养基后，用固定/变性液固定细胞同时使DNA变性。然后加入BrdU鼠源单抗检测掺入的BrdU（DNA变性使得掺入的BrdU易于为抗体所识别）。Anti-mouse IgG, HRP-linked antibody用来识别联胞检测抗体。加入HRP底物(TMB)进行显色。显色过程中吸光度的强度和掺入细胞的BrdU的量成正比, BrdU是细胞增殖的直接证明。卤代核苷酸如嘧啶类似物溴脱氧尿苷(BrdU)对于新生活细胞和组织中的DNA进行标记是有用的。BrdU掺入复制的DNA后代替胸苷，随后使用允许细胞周期中S期标记的细胞特异性单克隆抗体对BrdU进行免疫检测。溴脱氧尿苷脉冲标记细胞或组织后，BrdU (Bu20a)小鼠单克隆抗体可用于检测掺入单链DNA的BrdU。请参阅我们有关标记程序的详细步骤，以及应用于各种免疫检测的双链DNA变性程序(1-4)。

原厂资料：

Specificity / Sensitivity

BrdU Cell Proliferation Assay kit detects BrdU incorporation into cellular DNA during cell proliferation. The BrdU-labeled DNA has to be denatured to be detected by the BrdU Mouse mAb used in this kit. This BrdU Mouse mAb does not cross react with endogenous DNA. Depending on the cell type and the incubation time applied in the assay, 0.2-2x104 cells/well are sufficient for most experimental setups. For the best result, a cell number titration (Figure 1) is recommended.

Description

The BrdU Cell Proliferation Assay Kit detects 5-bromo-2’-deoxyuridine (BrdU) incorporated into cellular DNA during cell proliferation using an anti-BrdU antibody. When cells are cultured with labeling medium that contains BrdU, this pyrimidine analog is incorporated in place of thymidine into the newly synthesized DNA of proliferating cells. After removing labeling medium, cells are fixed and the DNA is denatured with our fixing/denaturing solution. Then a BrdU mouse mAb is added to detect the incorporated BrdU (The denaturing of DNA is necessary to improve the accessibility of the incorporated BrdU to the detection antibody). Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate TMB is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of BrdU incorporated into cells, which is a direct indication of cell proliferation.

Background

Halogenated nucleotides such as the pyrimidine analog bromodeoxyuridine (BrdU) are useful for labeling nascent DNA in living cells and tissues. BrdU becomes incorporated into replicating DNA in place of thymidine and subsequent immunodetection of BrdU using specific monoclonal antibodies allows labeling of cells in S phase of the cell cycle. After pulse-labeling cells or tissues with bromodeoxyuridine, BrdU (Bu20a) Mouse mAb can be used to detect BrdU incorporated into single stranded DNA. Please see our detailed protocol for information regarding the labeling procedure as well as denaturation of double stranded DNA for various immunodetection applications (1-4).

注意事项：

Note: This kit contains mixed storage components. Please store this entire kit at -20° C for long term storage. Upon first use, please allow components to thaw and then store each component as indicated on individual component labels.