BrdU Cell Proliferation Assay

Description

• General description: A non-isotopic enzyme immunoassay for the quantification of cell proliferation.Evaluation of cell cycle progression is essential for investigations in many scientific fields. Measurement of [3H] thymidine incorporation as cells enter S phase has long been the traditional method for the detection of cell proliferation. Subsequent quantification of [3H] thymidine is performed by scintillation counting or autoradiography. This technology is slow, labor intensive and has several limitations including the handling and disposal of radioisotopes and the necessity of expensive equipment. A well-established alternative to [3H] thymidine uptake has been demonstrated by numerous investigators. In these methods bromodeoxyuridine (BrdU), a thymidine analog replaces [3H] thymidine. BrdU is incorporated, into newly synthesized DNA strands of actively proliferating cells. Following partial denaturation of double stranded DNA, BrdU is detected immunochemically allowing the assessment of the population of cells, which are actively synthesizing DNA. The Calbiochem® BrdU Cell Proliferation Assay involves incorporation of BrdU into cells cultured in plates and BrdU immunolabeling using the cell layer as the solid phase. The resultant assay is sensitive, rapid, easy to perform and applicable to high sample throughput. In addition to evaluation of cell proliferation, information such as cell number, morphology and analysis of cellular antigens can be obtained from a single culture.
• Components: BrdU Label, Fixative/Denaturing Solution, BrdU Antibody, Antibody Diluent, Peroxidase Goat Anti-Mouse IgG, Conjugate Diluent, Substrate, Plate Wash Concentrate, Stop Solution, and a user protocol.
• Warning: Toxicity: Multiple Toxicity Values, refer to MSDS (O)
• Specifications: Assay Time: 3 h
• Principle: The Calbiochem® BrdU Cell Proliferation Assay is a non-isotopic immunoassay for the quantitation of bromodeoxyuridine incorporation into newly synthesized DNA of actively proliferating cells.
• Storage and Stability: Upon arrival store the entire contents of the kit at -20°C, in a non-frostfree freezer.Prior to use:1. Remove the Fixative/Denaturing Solution and place at room temperature for at least 4 h prior to use. Precipitates that may occur while cold should go back into solution.2. Reconstitute the Peroxidase Goat Anti-Mouse IgG with the appropriate volume of 1X PBS.• For the 200 test kit, reconstitute Peroxidase Goat Anti-Mouse IgG in 125 µl of 1X PBS. Use Conjugate Diluent for any further lot-specific dilution.• For the 1000 test kit, reconstitute Peroxidase Goat Anti-Mouse IgG in 250 µl of 1X PBS. Use Conjugate Diluent for any further lot-specific dilution.Allow solution to stand at room temperature for 10 min. (or until solution is clear). Divide into small aliquots; aliquots not to be used that same day should be stored at -20°C in a non-frostfree freezer.Once the kit components have been thawed for use:• The BrdU label and 100X Anti-BrdU Antibody should be divided into small aliquots and stored at -20°C in a non-frostfree freezer with the Peroxidase Goat Anti-Mouse IgG; avoid multiple freeze/thaw cycles.• All the other kit components (Substrate, Antibody Diluent, Conjugate Diluent, 20X Plate Wash Concentrate, Fixative/Denaturing Solution, Stop Solution) should be stored at 4°C.
• Other Notes: Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
• Legal Information: CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany