MilliporeSigma
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11647229001

Roche

Cell Proliferation ELISA, BrdU (colorimetric)

sufficient for ≤1,000 tests

Synonym(s):
cell proliferation

usage

sufficient for ≤1,000 tests

Quality Level

manufacturer/tradename

Roche

shipped in

wet ice

storage temp.

2-8°C

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.

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.

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 cytokines
  • Determination of the inhibitory or stimulatory effects of various compounds on cell proliferation in environmental and biomedical research, and in the food, cosmetic, and pharmaceutical industries
  • Measurement of the immunoreactivity of lymphocytes, stimulated by mitogens or antigens
  • Analysis of the chemosensitivity of tumor cells to different cytostatic drugs in medical research
  • Testing of biocompatibility of various scaffolds, employed in bone tissue engineering, for bone cell growth

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 solution

Dilute 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 solution

Dissolve Anti-BrdU-peroxidase in 1.1 ml double-dist. water for 10 minutes and mix thoroughly.

Anti-BrdU-peroxidase working solution

Dilute 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 solution

Washing solution

Dilute 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 solution
The 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 solution
At 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 solution
Prepare shortly before use. Do not store.
Washing solution
At 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

Other Notes

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

Kit Components Only

Product No.
Description

  • BrdU Labeling Reagent

  • FixDenat ready-to-use

  • Anti-BrdU-peroxidase antibody

  • Antibody Dilution Solution ready-to-use

  • Washing Buffer PBS 10x concentrated

  • Substrate Solution TMB ready-to-use

Pictograms

FlameExclamation mark

Signal Word

Danger

Hazard Statements

Hazard Classifications

Eye Irrit. 2 - Flam. Liq. 2 - Skin Sens. 1

Storage Class Code

3 - Flammable liquids

WGK

WGK 2

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Quotes and Ordering

Jia-Ang Shi et al.
International journal of molecular medicine, 34(1), 237-243 (2014-04-24)
It is known that microRNA-219 (miR-219) expression is downregulated in medulloblastoma. In the present study, we investigated the expression, targets and functional effects of miR-219 in D283-MED medulloblastoma cells. We first demonstrated that miR-219 not only inhibits proliferation, but also
Hongzhi Ma et al.
Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 36(8), 6103-6112 (2015-04-29)
Zinc finger protein, X-linked (ZFX) is a transcriptional factor involved in many physiological processes such as embryonic stem cell survival and self-renewal. Though ZFX dysfunctions have been identified in variant human diseases and especially in cancers, its pathological roles have
María Carolina Durán et al.
Journal of nanobiotechnology, 9, 47-47 (2011-10-22)
Reproducibly high transfection rates with low methodology-induced cytotoxic side effects are essential to attain the required effect on targeted cells when exogenous DNA is transfected. Different approaches and modifications such as the use of nanoparticles (NPs) are being evaluated to
Zsuzsanna E Toth et al.
Blood, 111(12), 5544-5552 (2008-02-13)
Granulocyte colony-stimulating factor (G-CSF) induces proliferation of bone marrow-derived cells. G-CSF is neuroprotective after experimental brain injury, but the mechanisms involved remain unclear. Stem cell factor (SCF) is a cytokine important for the survival and differentiation of hematopoietic stem cells.
Lutz Wollin et al.
The Journal of pharmacology and experimental therapeutics, 349(2), 209-220 (2014-02-22)
The tyrosine kinase inhibitor nintedanib (BIBF 1120) is in clinical development for the treatment of idiopathic pulmonary fibrosis. To explore its mode of action, nintedanib was tested in human lung fibroblasts and mouse models of lung fibrosis. Human lung fibroblasts

Articles

Cell Cycle Analysis Using a Nucleoside Triphosphate (NTP) Transporter Molecule for Rapid DNA Labeling in Living Cells

Regulation of the cell cycle involves processes crucial to the survival of a cell, including the detection and repair of genetic damage as well as the prevention of uncontrolled cell division associated with cancer. The cell cycle is a four-stage process in which the cell 1) increases in size (G1-stage), 2) copies its DNA (synthesis, S-stage), 3) prepares to divide (G2-stage), and 4) divides (mitosis, M-stage). Due to their anionic nature, nucleoside triphosphates (NTPs), the building blocks of both RNA and DNA, do not permeate cell membranes.

Cell Viability and Proliferation Assays

Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.

Protocols

Cell Proliferation ELISA, BrdU (colorimetric) Protocol & Troubleshooting

Cell Proliferation ELISA, BrdU (colorimetric) Protocol & Troubleshooting

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