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BrdU Cell Proliferation Assay

This proliferation assay is a non-isotopic immunoassay for quantification of BrdU incorporation into newly synthesized DNA of actively proliferating cells.

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Synonym(s):
Bromodeoxyuridine Assay

Quality Level

usage

sufficient for 1000 tests
sufficient for 200 tests

packaging

pkg of 1 96-well plate(s)

manufacturer/tradename

Calbiochem®

storage condition

OK to freeze
avoid repeated freeze/thaw cycles

input

sample type intact cells
sample type intact cells

detection method

colorimetric

shipped in

wet ice

storage temp.

−20°C

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This Item
116699150012750MAB3510
packaging

pkg of 1 96-well plate(s)

packaging

-

packaging

pkg of 200 assay(s)

packaging

-

manufacturer/tradename

Calbiochem®

manufacturer/tradename

Roche

manufacturer/tradename

Chemicon®

manufacturer/tradename

Chemicon®

storage condition

OK to freeze, avoid repeated freeze/thaw cycles

storage condition

-

storage condition

-

storage condition

-

input

sample type intact cells
sample type intact cells

input

-

input

-

input

-

detection method

colorimetric

detection method

chemiluminescent

detection method

colorimetric

detection method

-

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.
BrdU Cell Proliferation Assay is a non-isotopic immunoassay for quantification of BrdU incorporation into newly synthesized DNA of actively proliferating cells. It is sensitive, rapid, and easy to perform.
Note: 1 T = 1 test.
This proliferation assay is a non-isotopic immunoassay for quantification of BrdU incorporation into newly synthesized DNA of actively proliferating cells.

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.
Hardonk, M.J. and Harms, G. 1990. Acta histochemica, Suppl.39, 99.
Muir, D., et al. 1990. Analytical Biochemistry185, 377.
Lanier, T. L., et al. 1989. Carcinogenesis10, 1341.
Magaud, J.-P., et al. 1988. J. Immunol. Methods106, 95.
Collins, S.J. 1987. Blood70, 1233.
Porstmann, T., et al. 1985. J. Immunol. Methods82, 169.
Raza, A., et al. 1985. Cytometry6, 633.
Morstyn, G., et al. 1983. J. Clin. Invest.72, 1844.
Gratzner, H.G. 1982. Science218, 474.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Signal Word

Danger

Hazard Classifications

Carc. 2 - Eye Irrit. 2 - Flam. Liq. 2 - Met. Corr. 1 - Muta. 1B - Repr. 2 - Skin Irrit. 2 - Skin Sens. 1

Storage Class Code

3 - Flammable liquids

WGK

WGK 3

Flash Point(F)

69.8 °F

Flash Point(C)

21 °C


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