The BrdU cell proliferation assay is a non-isotopic immunoassay for the quantification of BrdU incorporation into newly synthesized DNA from actively proliferating cells. It is responsive, fast, and easy to do.
This proliferation assay is a non-isotopic immunoassay for the quantification of BrdU incorporation into newly synthesized DNA from actively proliferating cells.
A non-isotopic enzyme immunoassay for the quantification of cell proliferation. Assessment of cell cycle progression is essential for research in many scientific fields. Measurement of [3H] thymidine incorporation when cells enter S phase has long been the traditional method for detecting 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 handling and disposal of radioisotopes and the need for expensive equipment.
Numerous investigators have demonstrated a well-established alternative to [3H] thymidine uptake. In these methods, bromodeoxyuridine (BrdU), a thymidine analog, replaces [3H] thymidine. BrdU is incorporated into newly synthesized DNA strands of actively proliferating cells. After partial denaturation of the double-stranded DNA, BrdU is detected immunochemically, allowing the population of cells that actively synthesize DNA to be evaluated.
The Calbiochem BrdU cell proliferation assay involves the incorporation of BrdU into plated cells and immunostaining with BrdU using the cell layer as a solid phase. The resulting assay is sensitive, fast, easy to perform, and applicable to high throughput samples. In addition to assessing cell proliferation, information such as cell number, morphology, and analysis of cell 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-frost-free 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 the solution to stand at room temperature for 10 min. (or until a solution is clear). Divide into small aliquots; aliquots not to be used that same day should be stored at -20°C in a non-frost-free 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-frost free 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.
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