PrecisionAb Anti-BrdU Antibodies

BrdU Detection

When added to dividing cells, the thymidine analog 5-bromodeoxyuridine (BrdU) can be specifically incorporated into newly-synthesized DNA as 5-bromouracil (Figure 1). This incorporation happens during the S phase of the cell cycle, when cells are synthesizing new DNA. Subsequent detection of BrdU, using highly specific antibodies, enables researchers to identify dividing cells. 

As BrdU is a thymidine analog, it is essential to test anti-BrdU antibodies for cross-reactivity with thymidine. Binding of a BrdU antibody to thymidine could create unwanted background signal and affect experimental results. However, many anti-BrdU antibodies recognize the other halogenated thymidine analogs 5′-chloro-2′-deoxyuridine (CldU) and 5′-iodo-2′-deoxyuridine (IdU) (Liboska et al. 2012, Kimoto et al. 2008), due to the structural similarities of the three different halogenated thymidine analogs.

Knowing the cross-reactivity of your anti-BrdU antibody is critical when performing double-labeling experiments with BrdU and other nonhalogenated thymidine analogs, such as 5′-ethynyl-2′-deoxyuridine (EdU).


Fig. 1. BrdU (BrdU) can be specifically incorporated into newly-synthesized DNA as 5-bromouracil.


Fig. 1. BrdU can be specifically incorporated into newly-synthesized DNA as 5-bromouracil.


Extensively Validated Anti-BrdU Antibodies

Bio-Rad’s PrecisionAb Anti-BrdU Antibodies have been tested for cross-reactivity with thymidine and other thymidine analogs (Table 1). They have also been extensively tested for suitability in specific applications including flow cytometry and fluorescent immunocytochemistry.
 


Fig. 2.  A, Jurkat cells were treated with 100 μM BrdU and stained with Human Anti-BrdU Antibody (HCA320). B, HeLa cells were treated with 100 μM BrdU and stained with Human Anti-BrdU Antibody (HCA320).

Table 1. Cross-reactivity results for flow cytometry.

Cat#

BrdU

CIdU

EdU

IdU

Thymidine

HCA320

+

+

-

-

-

HCA321

+

+

-

-

-

HCA322

+

+

-

+

-

HCA323

+

+

-

-

-

MCA6143

+

+

+

+

-

MCA6144

+

+

-

-

-

Fig. 2. HeLa cells treated with BrdU and stained with Rat Anti-BrdU Antibody (MCA6144).

Fig. 3. HeLa cells treated with BrdU and stained with Rat Anti-BrdU Antibody (MCA6144).

Bio-Rad also has a range of rabbit, rat, and recombinant HuCAL® Anti-BrdU antibodies, suitable for the detection of BrdU in samples generated from mice, when combined with a suitable cross-adsorbed secondary antibody. Find out more on cross-adsorbed secondary antibodies.

Reduced Antibody Batch-to-Batch Variability with HuCAL Technology

As Bio-Rad’s HuCAL anti-BrdU antibodies are recombinant antibodies synthetically generated using the HuCAL technology, the sequences of the final antibodies are always known. This ensures batch-to-batch consistency and long-term security of supply. These antibodies are also chimeric, made by combining genetic material from rat or rabbit with human IgG. HCA320 and HCA321 have rabbit IgG chimeric regions, allowing them to be detected using an anti-rabbit IgG secondary antibody such as STAR36D488GA. HCA322 and HCA323 have rat IgG chimeric regions, allowing them to be detected using an anti-rat IgG antibody such as STAR16D488GA.

Visit the BrdU resources page, for the full range of anti-BrdU antibodies as well as protocols, tips and tricks, articles, and podcasts to help you with your BrdU labeling experiments.

PrecisionAb Anti-BrdU Antibodies

    DescriptionSpecificityTargetFormatHostIsotypeClone Applications Citations Product Type Code

    References:

    • Kimoto M et al. (2008). Label-retaining cells in the rat submandibular gland. J Histochem Cytochem 56, 15-24.
    • Liboska R et al. (2012). Most anti-BrdU antibodies react with 2′-deoxy-5′-ethynyluridine – the method for the effective suppression of this cross-reactivity. PLoS One 7, e51679.