Brdu incorporation assay kit

Manufactured by Cell Signaling Technology

Sourced in United States, Morocco, Japan

The BrdU incorporation assay kit is a laboratory tool used to measure cell proliferation. It detects the incorporation of the synthetic nucleoside bromodeoxyuridine (BrdU) into the DNA of actively dividing cells, providing a quantitative assessment of cell proliferation.

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Lab products found in correlation

Obatoclax, by Merck Group (1 mentions) Pd98059, by Merck Group (1 mentions) Microplate reader, by Bio-Rad (1 mentions) Phospho gsk 3β ser9, by Cell Signaling Technology (1 mentions) Elk 1, by Cell Signaling Technology (1 mentions) Pten sirna, by Santa Cruz Biotechnology (1 mentions) Control sirna, by Santa Cruz Biotechnology (1 mentions) Gsk3β, by Cell Signaling Technology (1 mentions) Phospho akt ser473, by Cell Signaling Technology (1 mentions) Phospho erk thr202 tyr204, by Cell Signaling Technology (1 mentions) Molecular biology grade reagents, by Merck Group (1 mentions) Cell counting kit 8 cck 8 assay kit, by Dojindo Laboratories (1 mentions) Tgf β1, by R&D Systems (1 mentions) Fibroblast growth factor 2 (fgf2), by R&D Systems (1 mentions)

Close spelling variants of this product

5-Bromo-2′-deoxyuridine (BrdU) incorporation assay kit BrdU incorporation BrdU assay kit BrdU assay

5 protocols using brdu incorporation assay kit

BrdU-based Cell Proliferation Assay

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Cell proliferation was evaluated by a BrdU incorporation assay kit (Cell Signaling Technology, USA) according to the manufacturer’s instructions. In brief, OECs and OEC-3Cs (5 × 10³ cells/well) were seeded in 96-well plates in complete EGM2 medium. The cells were then incorporated with BrdU solution for 24 h, and the relative cell proliferation level was determined by measuring the absorbance at 450 nm.

Kim Y.J., Ji S.T., Kim D.Y., Jung S.Y., Kang S., Park J.H., Jang W.B., Yun J., Ha J., Lee D.H, & Kwon S.M. (2018). Long-Term Priming by Three Small Molecules Is a Promising Strategy for Enhancing Late Endothelial Progenitor Cell Bioactivities. Molecules and Cells, 41(6), 582-590.

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Nicotine Impacts on Cell Proliferation

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BrdU incorporation assay was used to determine the effects of nicotine on cell proliferation. This assay was performed using a BrdU incorporation assay kit (Cell Signaling, Danvers, Massachusetts) as described previously.^{21 (link)} Briefly, 48 hours after nicotine treatment, mESCs were treated with 10 mM BrdU for 4 hours and incubated with anti-BrdU peroxidase conjugate for 90 minutes before adding a tetramethylbenzidine substrate. Incorporated BrdU was measured by an enzyme-linked immunosorbent assay reader, and data were represented as fold change compared to control group.

Qu Q., Zhang F., Zhang X, & Yin W. (2017). Bidirectional Regulation of Mouse Embryonic Stem Cell Proliferation by Nicotine Is Mediated Through Wnt Signaling Pathway. Dose-Response, 15(4), 1559325817739760.

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Quantifying Cell Proliferation using BrdU Assay

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The BrdU incorporation assay was performed as described previously (Lee and Lee, 2014 (link)). In brief, after 24 h of seeding with 2 × 10⁴ cells/ well in 96-well plates, the cells were treated with obatoclax or PD98059 (Sigma, USA). Cell proliferation was assayed using the BrdU incorporation assay kit (Cell Signaling Technology) according to the manufacturer’s instructions. BrdU was added 12 h before culture termination. At the end of culture, cells were fixed with fixing solution for 30 min at RT, rinsed twice with PBS, incubated with monoclonal anti-BrdU antibody for 1 h, followed by anti-mouse secondary antibody for 30 min. After the final wash, the substrate was added to the wells and then the stop solution was administered. The proportion of total cells incorporating BrdU into the nucleus was determined by reading the absorbance on a microplate reader (Bio-Rad) at 450 nm to calculate cell viability.

Oh J.H., Lee J.Y., Park J.H., No J.H, & Lee N.K. (2015). Obatoclax Regulates the Proliferation and Fusion of Osteoclast Precursors through the Inhibition of ERK Activation by RANKL. Molecules and Cells, 38(3), 279-284.

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Palbociclib and Ribociclib Cytotoxicity Assay

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Palbociclib and ribociclib were from Elleckchem. They were prepared to 10 mM with dimethyl sulfoxide (DMSO) as stock solution and stored at −20°C. Then they were freshly diluted with cell culture medium to certain concentrations. 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazol-iumbromide (MTT), antibody against β-actin (Cat# A2228), and monoclonal anti-HA-peroxidase antibody (Cat# H6533) were purchased from Sigma. Anti-Myc-peroxidase antibody (Cat# R951-25) was from Thermo Fisher. BrdU incorporation assay kit (#6831) and antibodies against PTEN (Cat# 9188), phospho-Rb (Ser280) (Cat# 8181), Rb (Cat# 9309), phospho-Akt (Ser473) (Cat# 4060), Akt (Cat# 4685), phospho-ERK (Thr202/Tyr204) (Cat# 4370), ERK (Cat# 4695), phospho-GSK-3β (Ser9) (Cat# 5558), GSK-3β (Cat# 12456), phospho-Elk-1 (Ser383) (Cat# 9186), and Elk-1 (Cat# 9182) were from Cell signaling (Beverly, MA). The horseradish peroxidase linked IgG secondary antibodies were obtained from GE Healthcare. Plasmids of constitutively active Akt (CA, Cat# 14751) (Scheid et al., 2002 (link)), ERK (CA, Cat# 39194) (Robinson et al., 1998 (link)), and PTEN (#78776) were from Addgene. PTEN siRNA (Cat# sc-29459) and control siRNA (Cat# sc-36369) were purchased from Santa Cruz. All chemicals and reagents not described above were from Sigma-Aldrich (St. Louis, MO).

Liu S., Yuan D., Li Y., Qi Q., Guo B., Yang S., Zhou J., Xu L., Chen T., Yang C., Liu J., Li B., Yao L, & Jiang W. (2019). Involvement of Phosphatase and Tensin Homolog in Cyclin-Dependent Kinase 4/6 Inhibitor-Induced Blockade of Glioblastoma. Frontiers in Pharmacology, 10, 1316.

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Quantifying Cell Responses to Growth Factors

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We used the recombinant mouse OPN, TGFβ1, and FGF2 supplied by R&D Systems (Minneapolis, MN, USA). Recombinant human bone morphogenetic protein 2 (BMP2) was purchased from CowellMedi (Busan, South Korea). Quantikine Mouse/Rat osteopontin, Mouse/Rat/Porcine/Canine TGFβ1, and Mouse/Rat FGF basic ELISA kits were obtained from R&D Systems. Cell Counting Kit-8 (CCK-8) assay kit was supplied by Dojindo Laboratories (Kumamoto, Japan), and BrdU incorporation assay kit was ordered through Cell Signaling Technology (Danvers, MA, USA). The molecular biology-grade reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA) unless stated otherwise.

Lee M.N., Hwang H.S., Oh S.H., Roshanzadeh A., Kim J.W., Song J.H., Kim E.S, & Koh J.T. (2018). Elevated extracellular calcium ions promote proliferation and migration of mesenchymal stem cells via increasing osteopontin expression. Experimental & Molecular Medicine, 50(11), 1-16.

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