Not available outside of the UK & Ireland.

Application

For research use only. Not for use in diagnostic procedures.The Cell Proliferation ELISA, BrdU (colorimetric) belongs to the second, improved generation of kits for measuring DNA synthesis. It is a precise, fast, and simple colorimetric alternative to quantitate cell proliferation based on the measurement of BrdU incorporation during DNA synthesis in replicating (cycling) cells. Thus, the Cell Proliferation ELISA can be used in many different in vitro cell systems. For example:Detection and quantification of cell proliferation induced by growth factors and cytokinesDetermination of the inhibitory or stimulatory effects of various compounds on cell proliferation in environmental and biomedical research, and in the food, cosmetic, and pharmaceutical industriesMeasurement of the immunoreactivity of lymphocytes, stimulated by mitogens or antigensAnalysis of the chemosensitivity of tumor cells to different cytostatic drugs in medical researchTesting of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth

General description

Colorimetric immunoassay for the quantification of cell proliferation, based on the measurement of BrdU incorporation during DNA synthesis:  a nonradioactive alternative to the [³H]-thymidine incorporation assay.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Packaging

1 kit containing 6 components.

Preparation Note

Working concentration: The kit antibody (Anti-BrdU-peroxidase) has, after reconstitution, a concentration of 7.5 U/ml, after dilution 0.075 U/ml. Instead of the kit antibody, you can also use the Anti-BrdU-Antibody, Fab fragments.This antibody is double-concentrated, so you have to dilute it after reconstitution in 1 ml with 1:200.Working solution: BrdU labeling solutionDilute BrdU labeling reagent 1:100 with sterile culture medium (resulting concentration: 100 µM BrdU).For one 96-well MP, 1 ml BrdU labeling solution is required if the cells were cultured in 100 µl /well (10 µl/well) and 2 ml BrdU labeling solution is required if the cells were cultured in 200 µl/well (20 µl/well).Anti-BrdU-peroxidase stock solutionDissolve Anti-BrdU-peroxidase in 1.1 ml double-dist. water for 10 minutes and mix thoroughly.Anti-BrdU-peroxidase working solutionDilute Anti-BrdU-peroxidase stock solution 1:100 with antibody dilution solution. For one 96-well MP dilute 100 µl Anti-BrdU-peroxidase stock solution in 10 ml antibody dilution solutionWashing solutionDilute Washing buffer concentrate 1:10 with double-dist. water.For one 96-well MP dilute 10 ml Washing buffer concentrate with 90 ml double-dist. water.Storage conditions (working solution): BrdU labeling solutionThe undiluted BrdU labeling reagent (1000x): At 2 to 8 °C for several months protected from light. The diluted BrdU labeling reagent: At 2 to 8 °C stable for several weeks. Store protected from light. For long-term storage it is recommended to store the BrdU labeling solution in aliquots at -15 to -25 °C.Anti-BrdU-peroxidase stock solutionAt 2 to 8 °C for several months. For long-term storage it is recommended to store the solution in aliquots at -15 to -25 °C.Anti-BrdU-peroxidase working solutionPrepare shortly before use. Do not store.Washing solutionAt 2 to 8 °C for several weeks.

Reconstitution

The working solution for the antibody should be phosphate buffered saline containing 1% BSA, pH 7.4

Specificity

The antibody conjugate reacts with the thymidine analogue 5-bromo-2′-deoxyuridine (BrdU) and with BrdU incorporated into DNA. For binding to BrdU incorporated into the DNA, the BrdU-labeled DNA has to be denatured. The antibody does not cross-react with any endogenous cellular components such as thymidine, uridine, or DNA.