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Safety Information

MAB4072

Sigma-Aldrich

Anti-BrdU Antibody, clone 131-14871

clone 131-14871, Chemicon®, from mouse

Synonym(s):

BrdU

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

mouse

Quality Level

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

131-14871, monoclonal

species reactivity (predicted by homology)

all

manufacturer/tradename

Chemicon®

technique(s)

flow cytometry: suitable
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable

isotype

IgG1

shipped in

wet ice

target post-translational modification

unmodified

General description

The thymidine analog 5-bromo-2′-deoxyuridine (BrdU) is a common reagent used for both cell proliferation assays and for the detection of apoptotic cells. BrdU is readily incorporated into newly synthesized DNA, labeling cells that have progressed through the S-phase of the cell cycle and thereby serving as a marker for proliferating cells.

Specificity

Recognizes BrdU (Bromodeoxyuridine).

Application

Anti-BrdU Antibody, clone 131-14871 is a Mouse Monoclonal Antibody for detection of Bromodeoxyuridine also known as BrdU & has been validated in FC, IHC, IHC(P). This bromodeoxyuridine antibody is expect to react in all species.
Immunohistochemistry on deparaffinized sections of kidney, duodenum, and liver tissue: 1:1000-1:1250.

Flow cytometry on 100μL of 106 cells: 1:5000.

Optimal dilutions must be determined by the end user.

IMMUNOHISTOCHEMICAL PROCEDURE

FOR PARAFFIN EMBEDDED TISSUE SECTIONS

STAINING PROCEDURE:

1. Deparaffinize sections in xylene 3 x 5 minutes. Resin embedded or GMA (Glycol methacrylate, 2-hydroxyethyl methacrylate) treated sections it is unnecessary to deparaffinize as there is no paraffin.

2. Place slides in graded alcohols (100, 95, 90%) to water at 2 minute intervals, and air dry. (Omit for GMA sections)

3. Circle sections with PAP pen or diamond pen to identify and dry thoroughly.

4. For GMA or resin sections, place slides in 0.5% Tween/PBS (pH 7.6) for 29 minutes.

5. Incubate in prewarmed (40°C) 1N HCl for 1 hour.

6. Wash 3 x 3 minutes with PBS (pH 7.6). Slides may be held at this point and the procedure continued the following day.

7. Incubate in 1X Trypsin solution (0.02-0.05% w/v, prewarmed to 40°C) for 20 minutes at room temperature for NBF (normal buffered formalin) fixed tissue or 10 minutes for Carnoy′s fixed tissue.

8. Wash 3 x 5 minutes with PBS (pH 7.6).

9. Incubate sections in 1% H2O2 (10 mL 30% H2O2 in 290 mL methanol) for 20 minutes, or GMA sections in 3% H2O2 (10 mL 30% H2O2 in 90 mL water) for 3 minutes.

10. Wash 2 x 2 minutes with PBS (pH 7.6).

11. Using the Vector ABC kit, add 3 drops of normal horse serum to 10 mL of PBS. Use this solution to block the slides for 20 minutes at room temperature.

12. Blot the slides of excess normal horse serum and incubate with MAB4072 diluted in PBS/BSA/Tween 20 for 1 hour at room temperature.

13. Wash 3 x 3 minutes with PBS (pH 7.6).

14. Add 3 drops of normal horse serum to 10 mL of PBS and then add 1 drop of biotinylated antibody stock. Incubate the slides with this diluted second antibody for 30 minutes at room temperature.

15. Prepare the Vectastain ABC Reagent by adding 2 drops of reagent A to 5 mL of PBS. Then add 2 drops of Reagent B to the same mixing bottle. Allow to sit for about 30 minutes prior to use.

16. Wash 3 x 3 minutes with PBS (pH 7.6).

17. Incubate the slides with ABC Reagent for 30 minutes at room temperature.

18. Wash 3 x 3 minutes with PBS (pH 7.6).

19. Prepare the substrate by mixing an equal volume of 0.02% H2O2 (made in distilled water from a 30% stock solution) and 0.1% DAB made in 0.1 M Tris buffer, pH 7.2. The H2O2 should be freshly prepared from concentrated stock. Because many peroxide substrates are unstable in the presence of H2O2 or when exposed to light, the substrate should be prepared just prior to use. Since DAB is a suspected carcinogen, care should be taken in handling and disposing of all peroxidase substrates.

20. Stain the slides with DAB substrate for 2-5 minutes.

21. Wash 2 x 2 minutes with distilled water.

22. Counterstain with hemotoxylin, dehydrate through xylene, and mount the coverslip.
Research Category
Epigenetics & Nuclear Function
Research Sub Category
Cell Cycle, DNA Replication & Repair

Physical form

Format: Purified
Liquid in 10 mM Phosphate buffer, pH 7.4 containing 150 mM NaCl and 0.1% sodium azide.
Protein A purified

Storage and Stability

Maintain for 2 years at -20°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Analysis Note

Control
Brdu treated cells

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

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

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Regulatory Listings

Regulatory Listings are mainly provided for chemical products. Only limited information can be provided here for non-chemical products. No entry means none of the components are listed. It is the user’s obligation to ensure the safe and legal use of the product.

JAN Code

MAB4072:


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Visualization of nascent transcripts on Drosophila polytene chromosomes using BrUTP incorporation.
W Y Chang et al.
BioTechniques, 29(5), 934-936 (2000-11-21)
Peng-Wei Tseng et al.
Frontiers in genetics, 13, 816955-816955 (2022-04-12)
Unlike gonochoristic fishes, sex is fixed after gonadal differentiation (primary sex determination), and sex can be altered in adults (secondary sex determination) of hermaphroditic fish species. The secondary sex determination of hermaphroditic fish has focused on the differences between testicular
Helge Rask-Andersen et al.
Hearing research, 203(1-2), 180-191 (2005-04-28)
Time lapse video recordings of cultured adult human and guinea pig spiral ganglion (hSG and gpSG) show that mitogen responsive progenitor/stem cells develop in the form of spheres that proliferate and differentiate into mature neurons and glia cells. Neurospheres, cultured
Ahmad Galuta et al.
Frontiers in neuroscience, 14, 607-607 (2020-07-07)
Improving the clinical translation of animal-based neural stem/progenitor cell (NSPC) therapies to humans requires an understanding of intrinsic human and animal cell characteristics. We report a novel in vitro method to assess spinal cord NSPCs from a small (rodent) and
Artem K Velichko et al.
Nucleic acids research, 43(13), 6309-6320 (2015-06-03)
Heat stress is one of the best-studied cellular stress factors; however, little is known about its delayed effects. Here, we demonstrate that heat stress induces p21-dependent cellular senescence-like cell cycle arrest. Notably, only early S-phase cells undergo such an arrest

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