Rat Anti-BrdU Antibody from Abcam

  • Product name - Anti-BrdU antibody [BU1/75 (ICR1)]
  • Description - Rat monoclonal [BU1/75 (ICR1)] to BrdU
  • Host species - Rat

Specificity

This antibody reacts with BrdU (5-Bromo-2’-deoxyuridine) linked to a protein carrier, free BrdU or BrdU in single-stranded DNA. It identifies nucleated cells that have had BrdU integrated into their DNA during S phase.

The antibody clone has been confirmed to react with EdU (5-Ethynyl-2’-deoxyuridine). In addition, it reacts with CldU (5-Chloro-2’-deoxyuridine) but with decreased staining. Some users have found that this antibody also reacts with IdU (5-Iodo-2’-deoxyuridine).

Tested applications

  • Appropriate for IHC-FoFr, ICC/IF, IHC-Fr, IHC-P, IHC-FrFl, Flow Cyt
  • Species reactivity - Not applicable
  • Immunogen - There are no particulars of the immunogen available for this antibody.

General notes

Protocol advice

This antibody detects single-stranded DNA, therefore, the DNA must be disentangled first. This can be carried out using DNAse; however, it doesn’t provide the best results. Based on the assay, heat denaturation or acid denaturation with 2M HCL is the most efficient. It is important to note that this step is crucial in any assay with this antibody and is the region that should be adjusted to optimize outcomes.

Unstained positive control slides from mice treated with BrdU (formalin-fixed, paraffin-embedded intestine sections) are available as BrdU control slides ab129956.

Abcam solely produces this monoclonal antibody:

Applications

The Abpromise guarantee from Abcam covers the use of ab6326 in the following tested applications. The application notes reports recommended starting dilutions; the end-user should determine the optimal dilutions/concentrations.

Application Abreviews Notes
ICC/IF ☆☆☆☆☆ 1/250.
IHC-FoFr ☆☆☆☆☆ 1/40.
IHC-P ☆☆☆☆☆ Use a concentration of 1 - 3 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-Fr ☆☆☆☆☆ 1/40 - 1/200. PubMed: 16670699In addition, found to work at 1/400. For PFA fixed tissue use at 1/40, from PMID 16373695.
Flow Cyt ☆☆☆☆☆ 1/25 - 1/200.
Rat IgG2a, kappa monoclonal [RTK2758] (Low endotoxin, Azide Free) (ab18450), is suitable for use as an isotype control with this antibody.
IHC-FrFl ☆☆☆☆☆ Use at an assay dependent concentration.

Target

Relevance

The immunocytochemical detection of bromodeoxyuridine (BrdU) integrated into DNA is a potent tool to explore the cytokinetics of neoplastic and normal cells. An accurate and inclusive technique to measure the degree of DNA-synthesis is in vivo or in vitro labeling of tumor cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies.

BrdU included into the newly produced DNA of S-phase cells may give an estimate for the fraction of cells in S-phase. Additionally, dynamic proliferative information like the S-phase transit rate and the potential doubling time can be yielded, with the help of bivariate BrdU/DNA flow cytometric analysis.

  • Cellular localization - Nuclear
  • Alternative names - BUdr antibody, Bromodeoxyuridine antibody