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BrdU Cell Proliferation

MBS396129

1000 Tests

650 USD

Assay Kit

BrdU Cell Proliferation

[BrdU Cell Proliferation]

BrdU Incorporated Cells

Store kit at -20 degree C until first use. Store at 4-8 degree C thereafter.

ELISA (EIA)

Intended Uses: The MyBioSource, Inc. BrdU Cell Proliferation Assay is a non-isotopic immunoassay for the quantitation of bromodeoxyuridine incorporation into newly synthesized DNA of actively proliferating cells.

Assay Kit; Proliferation

. Principle of the assay: Proliferation Assay involves incorporation of BrdU into cells cultured in microtiter plates using the cell layer as the solid phase. During the final 2 to 24 hours of culture BrdU is added to wells of the microtiter plate. BrdU will be incorporated into the DNA of dividing cells. To enable antibody binding to the incorporated BrdU cells must be fixed, permeabilized and the DNA denatured. This is all done in one step by treatment with Fixing Solution. Detector anti-BrdU monoclonal antibody is pipetted into the wells and allowed to incubate for one hour, during which time it binds to any incorporated BrdU. Unbound antibody is washed away and horseradish peroxidase-conjugated goat anti-mouse antibody is added, which binds to the Detector Antibody. The horseradish peroxidase catalyzes the conversion of the chromogenic substrate tetra-methylbenzidine (TMB) from a colorless solution to a blue solution (or yellow after the addition of stopping reagent), the intensity of which is proportional to the amount of incorporated BrdU in the cells. The colored reaction product is quantified using a spectrophotometer. Background: Evaluation of cell cycle progression is essentialfor 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 (1-8). 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. BrdU Cell Proliferation Assay involves incorporation of BrdU into cells cultured in microtiter plates 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.

1000 Tests

650 USD