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BrdU [5-Bromo-2'-deoxyuridine] *CAS 59-14-3*

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Physical properties
Molecular weight307.09
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC41116134

OverviewpdfSDSpdfProtocol


CAS
59-14-3
Molecular weight
307.09
5-Bromo-2-deoxyuridine (BrdU) is an analog of thymidine, and can be incorporated into the newly synthesized DNA of replicating cells (S-phase cells). BrdU is a commonly used in the detection of proliferating cells in living tissues. It works by substituting for thymidine during DNA replication and incorporating itself into the newly synthesized DNA (during S-phase). Antibodies specific for BrdU can then be used to detect the incorporated chemical, thus indicating cells that were actively replicating their DNA. Because BrdU can replace thymidine during the S phase of cell division, it can cause mutations.

Example protocol


PREPARATION OF STOCK SOLUTION

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

BrdU DMSO stock solution:
Make 1 - 10 mM DMSO stock solution by dissolving the BrdU in DMSO. Note: The DMSO stock solution is good for 6 months if stored at -20 °C.

PREPARATION OF WORKING SOLUTION

BrdU working solution:
Make 5-20 µM working solution in buffer of your choice. Note: BrdU working solution should be used promptly and be made fresh every time.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Pellet cells by centrifugation and resuspend the cells in completed growth media. Adherent cells in culture may be stained in situ on cover slips or in the cell culture wells.

  2. Add BrdU working solution and incubate it for 60 minutes to overnight as a guide. Note: The optimum incubation time will depend upon cell type and goal of the experiment.

  3. Fix cells with 70 - 80% alcohol or acid-ethanol for 20 - 30 minutes.

  4. Wash with PBS (3 times, 2 min. each).

  5. Proceed with immunocytochemical staining techniques.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of BrdU [5-Bromo-2'-deoxyuridine] *CAS 59-14-3* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM325.637 µL1.628 mL3.256 mL16.282 mL32.564 mL
5 mM65.127 µL325.637 µL651.275 µL3.256 mL6.513 mL
10 mM32.564 µL162.819 µL325.637 µL1.628 mL3.256 mL

Molarity calculator

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Citations


View all 2 citations: Citation Explorer
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Authors: Lee, Ching-Tien and Lin, Kun-Der and Hsieh, Cheng-Fang and Wang, Jiz-Yuh
Journal: Biomedicines (2023): 36
Heat-shock pre-treatment reduces liver injury and aids liver recovery after partial hepatectomy in mice
Authors: Oka, Yousuke and Akagi, Yoshito and Kinugasa, Tetsushi and Ishibashi, Nobuya and Iwakuma, Nobutaka and Shiratsuchi, Ichitarou and Shirouzu, Kazuo
Journal: Anticancer research (2013): 2887--2894

References


View all 55 references: Citation Explorer
DNA interstrand crosslinks are induced in cells prelabelled with 5-bromo-2'-deoxyuridine and exposed to UVC radiation
Authors: Wojcik A, Bochenek A, Lankoff A, Lisowska H, Padjas A, Szumiel I, von Sonntag C, Obe G.
Journal: J Photochem Photobiol B (2006): 15
Estrogen-induced region specific decrease in the density of 5-bromo-2-deoxyuridine-labeled cells in the olfactory bulb of adult female rats
Authors: Hoyk Z, Varga C, Parducz A.
Journal: Neuroscience (2006): 1919
Sequence-dependent formation of intrastrand crosslink products from the UVB irradiation of duplex DNA containing a 5-bromo-2'-deoxyuridine or 5-bromo-2'-deoxycytidine
Authors: Zeng Y, Wang Y.
Journal: Nucleic Acids Res. (2006)
Exposure to 5-bromo-2'-deoxyuridine induces oxidative stress and activator protein-1 DNA binding activity in the embryo
Authors: Sahambi SK, Hales BF.
Journal: Birth Defects Res A Clin Mol Teratol (2006): 580
The evaluation of early embryonic neurogenesis after exposure to the genotoxic agent 5-bromo-2'-deoxyuridine in mice
Authors: Kuwagata M, Ogawa T, Nagata T, Shioda S.
Journal: Neurotoxicology. (2006)
Flow cytometric method for enumeration and characterization of newly released polymorphonuclear leukocytes from the bone marrow using 5'-bromo-2'-deoxyuridine
Authors: Shih CH, Whalen BA, Goto Y, Hogg JC, van Eeden SF.
Journal: Am J Physiol Cell Physiol (2005): C757
5-Bromo-2'-deoxyuridine is selectively toxic to neuronal precursors in vitro
Authors: Caldwell MA, He X, Svendsen CN.
Journal: Eur J Neurosci (2005): 2965
Discrimination of cell nuclei in early S-phase, mid-to-late S-phase, and G(2)/M-phase by sequential administration of 5-bromo-2'-deoxyuridine and 5-chloro-2'-deoxyuridine
Authors: Yamada K, Semba R, Ding X, Ma N, Nagahama M.
Journal: J Histochem Cytochem (2005): 1365
Neuropathological examination of fetal rat brain in the 5-bromo-2'-deoxyuridine-induced neurodevelopmental disorder model
Authors: Ogawa T, Kuwagata M, Muneoka KT, Shioda S.
Journal: Congenit Anom (Kyoto) (2005): 14
Locomotor hyperactivity following prenatal exposure to 5-bromo-2'-deoxyuridine: neurochemical and behavioral evidence of dopaminergic and serotonergic alterations
Authors: Kuwagata M, Muneoka KT, Ogawa T, Takigawa M, Nagao T.
Journal: Toxicol Lett (2004): 63