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Brdu antibody

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The BrdU antibody is a laboratory reagent used to detect the incorporation of the nucleoside analog bromodeoxyuridine (BrdU) into cellular DNA. It is commonly used in cell proliferation and DNA synthesis studies.

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Lab products found in correlation

Bromodeoxyuridine (brdu), by Merck Group (3 mentions) 5 bromo 2 deoxyuridine brdu, by Merck Group (2 mentions) Eclipse 600, by Nikon (1 mentions) Tritc conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Evos cell imaging system, by Thermo Fisher Scientific (1 mentions) Bromodeoxyuridine (brdu), by BD (1 mentions) 5 fluorouridine furd, by Merck Group (1 mentions) Eclipse ti, by Nikon (1 mentions) Fitc conjugated secondary antibody, by Merck Group (1 mentions) Hoechst 33342, by Merck Group (1 mentions) β actin, by Merck Group (1 mentions) Brain derived neurotrophic factor (bdnf), by Abcam (1 mentions) Dab kit, by Zhongshan Golden Bridge Biotechnology (1 mentions) Protease inhibitor mixture, by Thermo Fisher Scientific (1 mentions) β actin antibody, by Santa Cruz Biotechnology (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) P75ntr antibody, by Abcam (1 mentions) Doublecortin dcx, by Abcam (1 mentions)

Spelling variants of this product

Anti-BrdU antibody (74 citations) Peroxidase-coupled anti-BrdU-antibody (32 citations) Mouse anti-BrdU monoclonal antibody (20 citations) Anti-BrdU primary antibody (4 citations) Mouse anti-BrdU primary antibody (3 citations) FITC‐coupled anti‐BrdU antibody (2 citations) Anti-BrdU primary antibody (B8434) (2 citations) Antibody against BrdU (2 citations) Mouse anti-BrdU-DNA antibody (2 citations) Mouse BrdU antibody (2 citations)

13 protocols using brdu antibody

1

BrdU Proliferation Assay Protocol

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After passage, the cells were seeded on cover slips which processed by 100 μg/mL poly-L-lysine and incubated with the proliferation marker 5-Bromo-2′-deoxyuridine (BrdU; 10 μmol/L, Sigma-Aldrich Inc. St. Louis, MO, USA) for 4 h. Before staining with 4′,6-diamidino-2-phenylindole (DAPI) and BrdU antibody (Sigma-Aldrich Inc. St. Louis, MO, USA) cells were fixed by 4% paraformaldehyde. Immunoreactive cells were visualized by fluorescence microscopy (Olympus, BX51, Japan) and a total of 5–7 randomly selected fields were captured. The minimal number of cells for counting was 200 in each condition, and data were collected from at least three independent experiments.
Lu Y., Lei S., Wang N., Lu P., Li W., Zheng J., Giri P.K., Lu H., Chen X., Zuo Z., Liu Y, & Zhang P. (2016). Protective Effect of Minocycline Against Ketamine-Induced Injury in Neural Stem Cell: Involvement of PI3K/Akt and Gsk-3 Beta Pathway. Frontiers in Molecular Neuroscience, 9, 135.
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2

BrdU Labeling for Cell Proliferation

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BrdU was injected into the pregnant mouse or newborn mouse (50 μg/g of body weight), 2 h prior to sacrificing the mouse. Samples were embedded as described previously. After hydration through graded ethanol, the sections were incubated in 0.1% PBST for 30 min and incubated in 2M HCl solution for 30 min under 37 °C, followed neutralization by boric acid for 10 min at RT. The sections were blocked with normal donkey serum at RT for 60 min before incubation in BrdU antibody (1:200, Sigma–Aldrich) overnight at 4 °C. Sections were washed in PBST three times. Second antibody donkey antimouse were diluted 1:500 for 60 min at RT.
Xie F., Zhu X., Liu X., Chen H, & Wang J. (2022). N6-methyladenosine (m6A) RNA methylation mediated by methyltransferase complex subunit WTAP regulates amelogenesis. The Journal of Biological Chemistry, 298(12), 102715.
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3

In vivo BrdU Labeling of Pancreatic Cells

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For in vivo BrdU labeling, mice were injected intraperitoneally with 50 mg/kg of BrdU in PBS, pH 7.6, and were sacrificed 1.5 h later. Pancreata were isolated and fixed in 10% formalin, embedded in paraffin, and processed by routine procedures. BrdU incorporation was detected by immunohistochemistry using BrdU antibody (Sigma) according to the manufacturer's instructions.
Song H.Y., Biancucci M., Kang H.J., O'Callaghan C., Park S.H., Principe D.R., Jiang H., Yan Y., Satchell K.F., Raparia K., Gius D, & Vassilopoulos A. (2016). SIRT2 deletion enhances KRAS-induced tumorigenesis in vivo by regulating K147 acetylation status. Oncotarget, 7(49), 80336-80349.
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4

BrdU Incorporation Immunofluorescence Protocol

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Cells in coating dishes were fixed with 2% paraformaldehyde after incubated with 3 μg/ml BrdU for 2 h. Incorporated BrdU was detected by immunofluorescence using a BrdU antibody (Sigma, St. Louis, MO, USA) according to manufacturer's instructions.
Dong Z., Huang M., Liu Z., Xie P., Dong Y., Wu X., Qu Z., Shen B., Huang X., Zhang T., Li J., Liu J., Yanase T., Zhou C, & Xu Y. (2016). Focused screening of mitochondrial metabolism reveals a crucial role for a tumor suppressor Hbp1 in ovarian reserve. Cell Death and Differentiation, 23(10), 1602-1614.
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5

BrdU Incorporation Assay for Cell Proliferation

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BrdU was added at the concentration of 50 μmol/L for 1 h. Cells were then fixed, permeabilized, and incubated with the BrdU antibody (1:1000, Sigma-Aldrich St. Louis, MO) followed by TRITC-conjugated secondary antibody (1:200, Santa Cruz), and finally stained with DAPI. A total of 100 cells were counted and photographed using Fluorescence microscopy (Nikon Eclipse 600) and percentages of positive cells were presented (mean ± SD).
Guo Y., Yuan X., Li K., Dai M., Zhang L., Wu Y., Sun C., Chen Y., Cheng G., Liu C., Strååt K., Kong F., Zhao S., Bjorkhölm M, & Xu D. (2019). GABPA is a master regulator of luminal identity and restrains aggressive diseases in bladder cancer. Cell Death and Differentiation, 27(6), 1862-1877.
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6

HUVEC Proliferation Assay by BrdU Incorporation

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Cell proliferation of HUVECs was determined by BrdU incorporation assay. HUVECs were seeded in 6-well culture plate and incubated with medium containing BrdU (3 μg/mL) and drugs for 24 h. The cells were then fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 for immunostaining with BrdU antibody (Sigma). Images were visualized and photographed using the EVOS Cell Imaging System (Thermo Fisher; Waltham, MA). Each treatment was performed in triplicate and at least 200 nuclei in each sample were counted for BrdU-positive cells.
Ai N., Chong C.M., Chen W., Hu Z., Su H., Chen G., Lei Wong Q.W, & Ge W. (2018). Ponatinib exerts anti-angiogenic effects in the zebrafish and human umbilical vein endothelial cells via blocking VEGFR signaling pathway. Oncotarget, 9(62), 31958-31970.
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7

Cell Proliferation Assay using BrdU Incorporation

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DNA synthesis was measured by BrdU incorporation into PC9 and A549 cells to determine cell proliferation. PC9 and A549 cells (2 × 103 cells/well) were seeded in a 96-well culture plate. After 48 h of culture, 10 μM BrdU (BD Pharmingen, USA) was added and the cells were incubated for another 2 h. The medium was removed and co-cultured with BrdU antibody (Sigma-Aldrich) for 60 min, followed by incubation with tetramethylbenzidine (peroxidase substrate) for 30 min. The absorbance of each well was determined at 450 nm.
Zhang Y., Xiao P, & Hu X. (2022). LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p. BMC Cancer, 22, 389.
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8

Investigating DDX49 Role in Pre-rRNA Transcription

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To study the role of DDX49 on the transcription of pre-ribosomal RNA, HeLa cells were seeded in the coverslips and transfected with control siRNA or DDX49 siRNA. After 48 h of transfection, the cells were pulse labeled with 2 mM of 5-fluorouridine (FUrd) (Sigma) for 20 min. After the incubation, the cells were fixed with 2% formaldehyde for 10 min and permeablized with 0.5% Triton X-100 for 10 min. The cells were then blocked with 1% BSA in PBST (PBS containing 0.1% Tween 20) for one hour and probed with BrdU antibody (Sigma, Cat. No. B8434) overnight at 4°C. After washing with PBS, the cells were incubated with FITC conjugated secondary antibody (Sigma, Cat. No. F2012) for 1 h at room temperature. After washing with PBS, the cells were treated with DAPI and embedded in Mowiol and images were taken in fluorescence microscope (Nikon Eclipse Ti) using 63× oil immersion objective.
Awasthi S., Verma M., Mahesh A., K. Khan M.I., Govindaraju G., Rajavelu A., Chavali P.L., Chavali S, & Dhayalan A. (2018). DDX49 is an RNA helicase that affects translation by regulating mRNA export and the levels of pre-ribosomal RNA. Nucleic Acids Research, 46(12), 6304-6317.
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9

Molecular Markers for Immune Cell Analysis

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DMBA, TPA, bromodeoxyuridine (BrdU), Hoechst 33342, and p38α antibody were purchased from Sigma (St. Louis, MO, USA). For immunofluorescence experiments, we used p38δ antibody obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). BrdU antibody was from Chemicon (Temecula, CA, USA). For immunoblot experiments, we used p38δ antibody purchased from the Division of Signal Transduction Therapy (Dundee, UK) [41 (link)]. Antibodies against CD45 and CD11b were from Thermo Fisher Scientific (Waltham, MA, USA), and CD3 antibody was from Rockland Immunochemicals (Limerick, PA, USA). Antibody against keratin 14 was obtained from Covance Research Products (Berkeley, CA, USA).
Kiss A., Koppel A.C., Murphy E., Sall M., Barlas M., Kissling G, & Efimova T. (2019). Cell Type-Specific p38δ Targeting Reveals a Context-, Stage-, and Sex-Dependent Regulation of Skin Carcinogenesis. International Journal of Molecular Sciences, 20(7), 1532.
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10

Quantifying Oligodendrocyte Progenitor Proliferation

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To identify the proliferation of oligodendrocyte progenitor cells (OPCs), mice were injected intraperitoneally with 200 mg/kg bromodeoxyuridine (BrdU; Sigma) at 4 and 2 h before killing (Armstrong et al. 2002 (link)). Brain sections from BrdU-injected mice were treated to permeabilize the tissue and denature the DNA, and were then incubated overnight with the following primary antibodies: rabbit polyclonal anti-NG2 antibody (1 : 200, Chemicon) and mouse monoclonal anti-BrdU antibody (1 : 50). The secondary antibodies (1 : 500 dilutions) used were Alexa 488-labeled goat anti-mouse antibodies for BrdU detection and Alexa 594-labeled goat anti-rabbit antibodies for NG2 detection. The sections were then washed and mounted to be imaged. For BrdU counting, three coronal sections per animal were analyzed with five randomly selected images within each area of interest (CC).The total numbers of all BrdU+ cells and BrdU+ NG2+ cells (oligodendrocyte progenitors) per field were counted.
Liu J., Tian D., Murugan M., Eyo U.B., Dreyfus C.F., Wang W, & Wu L.J. (2015). Microglial Hv1 proton channel promotes cuprizone-induced demyelination through oxidative damage. Journal of neurochemistry, 135(2), 347-356.
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