| 货号 | 5492S |
| 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. 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.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。 |
| 反应种属 | All |
| 应用 | FUNC |
| 目标/特异性 | 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. |
| 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 and denaturation of double stranded DNA for various immunodetection applications (1-4).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）。 |
| 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.将C2C12细胞以各种浓度接种到96孔板中进行无血清培养基培养过夜。随后加入各种不同浓度的血清并将细胞培养24小时。最后，加入10 μM BrdU孵育4小时。 | |
Figure 2. Treatment of MCF 10A cells with Human Epidermal Growth Factor (hEGF) #8916 increases cell proliferation as detected by the BrdU Cell Proliferation Chemiluminescent Assay Kit #5492. 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.用人表皮生长因子（hEGF）#8916处理MCF10A细胞可以促进细胞增殖，该过程经BrdU细胞增殖化学发光检测试剂盒#5492检测证实。将MCF10A细胞在96孔板中以1x104 cells/well的密度铺板，培养过夜。过夜时使用无血清培养基饥饿处理。随后加入hEGF培养24小时。最后，加入10 μM BrdU孵育4小时。 | |
Figure 3. Jurkat cells were seeded at 5x104 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.Jurkat细胞在96孔板中以5x104 cells/well的密度铺板，培养过夜。加入不同浓度的阿霉素处理细胞2小时。最后，加入10 μM BrdU孵育4小时。 |
