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BrdU (Bromodeoxyuridine / 5-bromo-2'-deoxyuridine) is an analog of the nucleoside thymidine used in the BrdU assay to identify proliferating cells.
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BrdU labeling can be performed in vitro for cell lines and primary cell cultures, or in vivo for labeling cells within a living animal. During the BrdU assay, BrdU is incorporated into replicating DNA and can be detected using anti-BrdU antibodies.
After BrdU labeling, an additional DNA hydrolysis step (sometimes referred to as a DNA denaturing step) may be required after fixation and permeabilization to allow the anti-BrdU antibody access to the BrdU within the DNA.
We recommend using our ab6326 BrdU antibody. This product has been cited in hundreds of publications and independently reviewed by our customers with an average 5-star rating. The clone and its conjugates are manufactured in-house.
For rapid, convenient results, we also recommend our BrdU staining IHC kit which contains all the reagents you need for staining cells or tissues which have incorporated BrdU. Or consider a BrdU ELISA kit for a quantitative measure of BrdU incorporation, without imaging.
EdU staining is an alternative method with a shorter, simpler protocol than that required for BrdU.
There are several methods for labeling cells in vivo with BrdU. Two commonly used methods are intraperitoneal injection and oral administration.
Note: BrdU incorporation into rapidly dividing tissues, such as the small intestine, will be detectable as soon as 30 minutes post-injection. However, for most tissue, you may need to wait up to 24 hours. The exact treatment time and dosage will need to be optimized for your tissue of interest.
Oral administration of BrdU is a non-invasive procedure and therefore useful for extended BrdU administration, although it may introduce variability into experiments due to lack of control over an animal’s water consumption.
Note: for mice, 225 mg/kg per day of BrdU (calculated by measuring water consumption volumes per animal) should achieve sufficient BrdU labeling. However, the exact dose should be optimized for individual experimental conditions.
Sodium borate buffer: 3.8g sodium borate (MW=381.4) + 100 ml distilled water. Adjust pH with NaOH.
Note: if using paraffin-embedded sections, ensure they are de-waxed before proceeding. Read our deparaffinization protocol here.
Note: some researchers have reported that they don’t perform the HCl hydrolysis step and simply perform heat-induced epitope retrieval before continuing with immunostaining.
BrdU can be used in conjunction with other antibodies to identify proliferating and newly differentiated neurons. See below for our suggestions.
A cellular marker for proliferation, the Ki67 protein is present in cells at cycle phases G1, S, G2 and M, but absent in resting (G0) cells. Ki67 antibodies can be used instead of, or in conjunction with, BrdU to label proliferating neurons.
A microtubule-associated phosphoprotein expressed by immature neurons. Doublecortin antibodies can be used in conjunction with BrdU to identify immature post-mitotic neurons.
A marker of mature neurons that can be used in conjunction with BrdU staining to identify newly differentiated neurons.
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