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BrdU [5-Bromo-2'-deoxyuridine] *CAS 59-14-3*
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
- 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.
- 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.
- Fix cells with 70 - 80% alcohol or acid-ethanol for 20 - 30 minutes.
- Wash with PBS (3 times, 2 min. each).
- 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 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 325.637 µL | 1.628 mL | 3.256 mL | 16.282 mL | 32.564 mL |
| 5 mM | 65.127 µL | 325.637 µL | 651.275 µL | 3.256 mL | 6.513 mL |
| 10 mM | 32.564 µL | 162.819 µL | 325.637 µL | 1.628 mL | 3.256 mL |
Molarity calculator
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Citations
View all 2 citations: Citation Explorer
SGLT2 Inhibitor Canagliflozin Alleviates High Glucose-Induced Inflammatory Toxicity in BV-2 Microglia
Authors: Lee, Ching-Tien and Lin, Kun-Der and Hsieh, Cheng-Fang and Wang, Jiz-Yuh
Journal: Biomedicines (2023): 36
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
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
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
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)
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
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)
Authors: Kuwagata M, Ogawa T, Nagata T, Shioda S.
Journal: Neurotoxicology. (2006)
Page updated on November 23, 2024
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