| 货号 | 6813S |
| 描述 | 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.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。 |
| 反应种属 | All |
| 应用 | W |
| 目标/特异性 | 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. |
| 供应商 | CST |
| 背景 | 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).卤代核苷酸如嘧啶类似物溴脱氧尿苷(BrdU)对活细胞和组织中的DNA进行标记是有用的。BrdU掺入复制的DNA后代替胸苷，随后使用标记细胞周期中S期细胞的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)。 |
| 存放说明 | -20C |
| 参考文献 | Darzynkiewicz, Z. and Juan, G. (2001) Curr Protoc Cytom Chapter 7, Unit 7.7. Leif, R.C. et al. (2004) Cytometry A 58, 45-52. Staszkiewicz, J. et al. (2009) Biochem Biophys Res Commun 378, 539-44. Rothaeusler, K. and Baumgarth, N. (2007) Curr Protoc Cytom Chapter 7, Unit7.31. |
Figure 1. C2C12 cells were seeded at varying density in serum free medium in a 96-well plate and incubated overnight. Serum was added to the plate at various concentrations and cells were incubated for 24 hr. Finally, 10 μM BrdU was added to the plate and cells were incubated for 4 hr. | |
Figure 2. Treatment of MCF 10A cells with Human Epidermal Growth Factor (hEGF) #8916 increases cell proliferation as detected by BrdU Cell Proliferation Assay Kit #6813. MCF 10A cells were seeded at 1x104 cells/well in a 96-well plate and incubated overnight. Cells were then starved in serum free medium overnight. hEGF was added to the plate and cells were incubated for 24 hr. Finally, 10 μM BrdU was added to the plate and cells were incubated for 4 hr. | |
Figure 3. Jurkat cells were seeded at 4x104 cells/well in a 96-well plate and incubated overnight. Cells were then treated with various concentrations of doxorubicin for 2 hr. Finally, 10 μM BrdU was added to the plate and cells were incubated for 4 hr. |
