BrdU Cell Proliferation Chemiluminescent Assay Kit

产品编号：CST-5492S 品牌：CST 原厂货号：5492S
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：

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】

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

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

Stable Peroxide Buffer【包装：25 ml，，运保温度：-20° C】

描述：

BrdU细胞增殖化学发光检测试剂盒能够检测在细胞增殖时掺入细胞DNA的BrdU。BrdU标记的DNA必须经过变性才能使用试剂盒中的BrdU Mouse mAb检测。BrdU Mouse mAb与内源性DNA没有交叉反应。0.2-2x104 cells/well足以应对大多数实验类型，细胞数目取决于细胞类型和孵育时间。为了得到最好的结果，建议对细胞进行梯度稀释（图1）。BrdU细胞增殖化学发光检测试剂盒使用Anti-BrdU抗体检测细胞增殖时掺入DNA中的5-bromo-2’-deoxyuridine (BrdU)。使用含有BrdU的培养基培养细胞，这种嘧啶类似物能够在新DNA和成时取代胸腺嘧啶。移除培养基后，使用我们的固定/变性液以固定细胞，变性DNA。DNA变性能提高BrdU的掺入以便被抗体检测到。然后加入BrdU鼠单抗以检测掺入的BrdU。Anti-mouse IgG，HRP-偶联的抗体用以结合抗体。然后加入化学发光试剂。发射光的强度以相对光单位测得（RLU），与掺入细胞的BrdU量成正比，是细胞增殖的直接指示。嘧啶类似物bromodeoxyuridine (BrdU)之类的卤化核苷酸能够用于标记活细胞和组织中的新生DNA。DNA复制时BrdU替代胸腺嘧啶掺入新生DNA，随后使用特异性的单抗对BrdU进行棉衣检测能够标记处于S期的细胞。使用溴脱氧尿苷对细胞或组织进行脉冲标记实验，BrdU(Bu20a)鼠单抗可以检测到单链DNA中掺入的BrdU。请查询我们的实验步骤获得各种免疫检测应用中标记和双链DNA变性的实验过程的具体信息（1-4）。

原厂资料：

Specificity / Sensitivity

BrdU Cell Proliferation Chemiluminescent Assay Kit detects BrdU incorporation into cellular DNA during cell proliferation. The BrdU-labeled DNA must 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 best results, 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. Denaturing of DNA is necessary to improve the accessibility of the incorporated BrdU to the detection antibody. A BrdU mouse mAb is then added to detect the incorporated BrdU. Anti-mouse IgG, HRP-linked Antibody is used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), 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 and 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.