Anti brdu antibody

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-BrdU antibody is a laboratory tool used to detect the incorporation of 5-bromo-2'-deoxyuridine (BrdU) into cellular DNA. BrdU is a synthetic thymidine analog that is incorporated into the DNA of proliferating cells during the S-phase of the cell cycle. The Anti-BrdU antibody specifically binds to BrdU, allowing researchers to visualize and quantify cellular proliferation using techniques such as immunohistochemistry or flow cytometry.

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Lab products found in correlation

Bromodeoxyuridine (brdu), by Merck Group (6 mentions) Anti rabbit cy3, by Jackson ImmunoResearch (2 mentions) Phospho histone h3, by Cell Signaling Technology (2 mentions) Lysozyme, by Agilent Technologies (2 mentions) Mmp 7, by R&D Systems (2 mentions) Target retrieval solution, by Agilent Technologies (1 mentions) Envision detection kit, by Agilent Technologies (1 mentions) Mtt assay, by Roche (1 mentions) Goat anti mouse igg fitc, by Santa Cruz Biotechnology (1 mentions) Facscalibur, by BD (1 mentions) Mtt cell proliferation assay kit, by American Type Culture Collection (1 mentions) Hoechst, by Merck Group (1 mentions) Apo one homogeneous caspase 3 7 assay, by Promega (1 mentions) Texas red, by Jackson ImmunoResearch (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Avantor (1 mentions) Facsaria 2, by BD (1 mentions) Brdu solution, by Merck Group (1 mentions) Mvx10 microscope, by Olympus (1 mentions)

Spelling variants of this product

Anti-BrdU (14 citations) BrdU antibody (4 citations) Anti-BrdU monoclonal antibody (4 citations) Mouse anti-BrdU antibody (3 citations) Monoclonal primary anti-BrdU antibody (2 citations)

19 protocols using anti brdu antibody

Immunohistochemical Analysis of Hippocampal Neurogenesis in Hypoxic Rats

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The hippocampi of hypoxic-ischemic rats
were removed and fixed in 4% paraformaldehyd
prior to paraffin embedding and sectioning. Following
dewaxing and rehydration, hippocampus
sections were stained with hematoxylin and
eosin (H&E) and immunohistochemistry performed.
To detect BrdU positive cells, paraffin
sections were retrieved using Target Retrieval
Solution (Dako, Carpinteria, USA). After endogenous
peroxidase and nonspecific protein
blocking, primary antibodies were incubated
overnight at 4˚C with anti-BrdU antibody (1:50,
Santa Cruz, USA) and rabbit control immunoglobulin
G (IgG). After washing, the immunoreactivity
of the sections was analyzed using
an EnVision detection kit (Dako, Denmark),
according to the manufacturer’s instructions.
Positive staining was brown.

Yu Q., Liu L., Lin J., Wang Y., Xuan X., Guo Y, & Hu S. (2015). SDF-1α/CXCR4 Axis Mediates The Migration of Mesenchymal Stem Cells to The Hypoxic-Ischemic Brain Lesion in A Rat Model. Cell Journal (Yakhteh), 16(4), 440-447.

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Cell Proliferation Assays for Growth Curves

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For growth curves, all cell lines were plated and grown under normal conditions for 1–4 days. Every day, cells were collected, diluted in Trypan blue to assess viability, and counted in the Burker’s chamber. Cell proliferation was further assessed by MTT assay according to the manufacturer's recommendations (Roche Diagnostics GmbH, Mannheim, Germany): 40 000 cells were seeded in triplicates in a 96-wells; 5 mg/ml MTT per well was added both 24 and 48 h after seeding. Cells were then incubated for 4 h and DMSO was used to dissolve crystals. 5-Bromo-2'-Deoxyuridine (BrdU) incorporation was also used to confirm cellular proliferation. Cells were plated and grown under normal conditions for 24 h. Cells were labeled with 3 μg/ml BrdU for 4 h, washed twice with PBS, fixed in 1% formaldehyde and then spotted on slides. immunofluorescence was performed using anti-BrdU antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), according to the manufacturer’s protocol.

Farina F.M., Inguscio A., Kunderfranco P., Cortesi A., Elia L, & Quintavalle M. (2017). MicroRNA-26a/cyclin-dependent kinase 5 axis controls proliferation, apoptosis and in vivo tumor growth of diffuse large B-cell lymphoma cell lines. Cell Death & Disease, 8(6), e2890-.

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Cell Proliferation Measurement by BrdU and PI

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Cell proliferation was measured by labeling cells with bromodeoxyuridine (BrdU) and propidium iodide (PI) prior to flow cytometry. BrdU-positive cells and PI staining were used to identify cells in S phase and expression of total DNA [25 (link), 26 (link)]. Cells were treated with SH003 for 48 h and labeled with 10 μM BrdU (Sigma-Aldrich) for 1 h before harvesting. Cells were then trypsinized and fixed in 70% ethanol on ice for 20 min. Next, cells were incubated with 2 M HCl/0.5% Tween-20/phosphate-buffered saline (PBS) for 30 min at room temperature. After washing with 1% bovine serum albumin (BSA) in PBS, cells were stained with anti-BrdU antibody (1:50; Santa Cruz, CA, USA) in buffer (0.5% Tween-20/1% BSA in PBS) for 30 min at room temperature. Cells were washed and then incubated for 30 min at room temperature with goat anti-mouse IgG-FITC (1:100; Santa Cruz). Washed cells were resuspended in PI for 30 min on ice. Cell proliferation was analyzed by FACSCalibur using CellQuest Pro software.

Choi Y.J., Choi Y.K., Lee K.M., Cho S.G., Kang S.Y, & Ko S.G. (2016). SH003 induces apoptosis of DU145 prostate cancer cells by inhibiting ERK-involved pathway. BMC Complementary and Alternative Medicine, 16, 507.

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FACS Analysis of Cell Cycle

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For FACS, cells were fixed with ice-cold 70% ethanol at −20 °C, and washed with phosphate-buffered saline (PBS), DNA was stained with a solution containing propidium iodide (5 µg/ml) and RNAse A (0.5 mg/ml) in PBS. For bivariate FACS analysis, cells were pre-labelled with 10 μM BrdU (Sigma) for 30 min, harvested via trypsinisation and fixed as describe above. Fixed cells were washed once with PBS and incubated with 2 M HCl, 0.5% (vol/vol) Triton X-100 for 30 min, with gentle mixing. After two washes in PBS, cells were incubated 2 h at room temperature with anti-BrdU antibody (Santa Cruz Biotechnology Inc.) and diluted 1/30 in PBS containing 1% BSA, 0.5% Tween 20. Then, BrdU staining was revealed with Alexa Fluor 488-conjugated goat anti-mouse antibody and DNA with a 7AAD (1.5 µg/ml) and RNAse A (0.5 mg/ml) in PBS. Finally, cells were analysed on a FACS Calibur (BD Biosciences) using Cell Quest Pro and Flow Jo software.

Charrasse S., Gharbi-Ayachi A., Burgess A., Vera J., Hached K., Raynaud P., Schwob E., Lorca T, & Castro A. (2017). Ensa controls S-phase length by modulating Treslin levels. Nature Communications, 8, 206.

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Cell Proliferation Assays by BrdU and MTT

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Cell proliferation was identified by 5-bromo-2′-deoxyuridine (BrdU) incorporation and MTT method. Briefly, cells were plated into 6-well glass slides and incubated with 30 µM BrdU (Sigma–Aldrich, St. Louis, MO) in culture medium for 4 h. The cells were then treated with a mouse monoclonal anti-BrdU antibody (1:100, Santa Cruz Biotechnology, Inc., Santa Cruz, CA), followed by a 1:1,000 dilution of the secondary antibody with Alexa Fluo^® 488 green (Molecular Probes, Eugene, OR). For nucleus staining, the cells were incubated with a 1:5,000 dilution of Hoechst (Sigma–Aldrich). Images were obtained in a Nikon inverted microscope. Cell proliferation was expressed as a percentage of the number of BrdU positive cells relative to total number of Hoechst positive nuclei. For MTT assay, cells were seeded into 96-well plates with 1.0 × 10³ cells/well and detected at days 1–6, respectively, using MTT cell proliferation assay kit (ATCC, Manassas, VA).

Wu L., Wang F., Donly K.J., Wan C., Luo D., Harris S.E., Macdougall M, & Chen S. (2015). Establishment of Immortalized Mouse Bmp2 Knock-Out Dental Papilla Mesenchymal Cells Necessary for Study of Odontoblastic Differentiation and Odontogenesis. Journal of cellular physiology, 230(11), 2588-2595.

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Cellular Senescence and HIV Protein Impacts

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Cellular senescence was evaluated by the PDL value calculated as log2 (D5/D0), where D0 and D5 are the number of cells at seeding and harvesting, respectively (Hernandez-Vallejo et al., 2013 (link)). Dividing cells were identified by measuring bromodeoxyuridine (BrdU) incorporation (BD Biosciences Pharmingen, San Diego, CA, USA). Upon day 9 and day 19 post-HIV-protein treatment, cells were incubated for 24 h with BrdU (15 μmol/L), then fixed and permeabilized. Anti-BrdU antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was revealed using secondary antibodies coupled to Texas Red (Jackson ImmunoResearch Laboratories, West Grove, PA, USA). Cell nuclei were visualized after diamidino-phenylindole hydrochloride staining (DAPI, Sigma-Aldrich). Dividing cells, examined by fluorescence microscopy, were counted in four randomly selected fields and expressed as a percentage of total cells. The percentage of apoptotic cells was determined using Apo-ONE homogeneous Caspase-3/7 assay (Promega Biosciences, San Luis Obispo, CA, USA). Quantification was performed on a plate fluorescence reader (Spectrafluor Plus, Tecan-France, Trappes, France) at 530 nm.

Beaupere C., Garcia M., Larghero J., Fève B., Capeau J, & Lagathu C. (2015). The HIV proteins Tat and Nef promote human bone marrow mesenchymal stem cell senescence and alter osteoblastic differentiation. Aging Cell, 14(4), 534-546.

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Immunolabeling Protocols for Neural Cell Types

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Chemical reagents were purchased from the following sources: mouse monoclonal anti-BrdU antibody from Santa Cruz Biotechnology (Dallas, TX, USA), ProLong Gold anti-fade reagent, Alexa Fluor 488 goat anti-rabbit IgG (H+L) antibody (Catalog No. A11008) and Alexa Fluor 555 goat anti-mouse IgG (H+L) antibody (Catalog No. A21424) from Life Technologies (Carlsbad, CA, USA); anti-doublecortin (DCX) (Catalog No. ab18723) and anti-NeuN (Catalog No. ab177487) antibodies, and chicken polyclonal anti-glial fibrillary acidic protein (GFAP) antibodies from Abcam (Cambridge, MA, USA); anti-tyrosine hydroxylase sheep polyclonal antibody (Catalog No. AB1542) from Millipore (Burlington, MA, USA); anti-GAD67 mouse monoclonal antibody (Catalog No. ab26116) from Abcam (Branford, CT, USA); paraformaldehyde (PFA) from ACROS Organics (Morris Plains, NJ, USA); bovine serum albumin (BSA) from AMRESCO (Solon, OH, USA); and normal goat serum and normal donkey serum from Jackson Immuno Research Labs (West Grove, PA, USA). All reagents were of analytical grade, HPLC grade, or the best available pharmaceutical grade.

Adamson S.X., Zheng W., Agim Z.S., Du S., Fleming S., Shannahan J, & Cannon J. (2021). Systemic Copper Disorders Influence the Olfactory Function in Adult Rats: Roles of Altered Adult Neurogenesis and Neurochemical Imbalance. Biomolecules, 11(9), 1315.

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Evaluating miR-455-3p Effects on Proliferation

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To determine the effects of miR-455-3p mimics or inhibitors on cell proliferation, we performed BrdU assay according to the manufacturer’s instructions. Briefly, the cells were seeded in 96-well plates (2×10³ cells/well) and were transfected with miR-455-3p mimics or inhibitors. After 48 h of transfection, 10 μM BrdU solution (Sigma-Aldrich, St. Louis, MO) was added to each plate and incubated at 37°C for an additional 2 h. The cells were washed 3 times, resuspended in a mixture of washing buffer, and incubated with 4 M hydrogen chloride (HCl) for 30 min at room temperature. Thereafter, the cells were resuspended in Borax buffer, washed, re-suspended in washing buffer, and then labeled with anti-BrdU antibody (Santa Cruz Biotechnology) for 1 h at 4°C in the dark. The cells were then incubated with secondary green-fluorescence dye conjugated antibody (Santa Cruz Biotechnology) for 45 min at room temperature. Finally, the cells were subjected to flow cytometric (FCM) analysis. Absorbance at 488 nm was recorded on a FACS Aria II device (BD Biosciences).

Zheng J., Lin Z., Zhang L, & Chen H. (2016). MicroRNA-455-3p Inhibits Tumor Cell Proliferation and Induces Apoptosis in HCT116 Human Colon Cancer Cells. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 22, 4431-4437.

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Quantifying Cell Proliferation in Nanofibrous Scaffolds

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The cells were cultured for 4 hours in the presence of 10 μM BrdU (Sigma-Aldrich), fixed in 4% paraformaldehyde for 15 minutes at room temperature, and incubated with blocking buffer (PBS containing 10% normal goat serum, 0.3% Triton X-100, and 100 ng/mL RNase A) for 60 minutes at room temperature. The cells were then washed in PBS, incubated with 1 N HCl for 30 minutes at room temperature, and washed in Hanks’ Balanced Salt Solution followed by PBS at room temperature. After overnight incubation at 4°C with anti-BrdU antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA) in blocking buffer, the cells were washed in PBS and incubated with fluorescent-conjugated secondary antibody (Alexa Fluor 488-goat anti-mouse, 1:500) for 1 hour at room temperature. The cell nuclei were counterstained with DAPI.
Fluorescent images of the nanofibrous scaffolds were recorded using an Axiovert 100 LSM710 laser scanning confocal microscope (Carl Zeiss, Oberkochen, Germany). The number of live and dead cells (showing green and red fluorescence, respectively) and amounts of BrdU and Ki-67 protein staining were counted in five fields at a magnification of 200×, and the percentages of live cells and positive cells were calculated.

Chen J., Yan C., Zhu M., Yao Q., Shao C., Lu W., Wang J., Mo X., Gu P., Fu Y, & Fan X. (2015). Electrospun nanofibrous SF/P(LLA-CL) membrane: a potential substratum for endothelial keratoplasty. International Journal of Nanomedicine, 10, 3337-3350.

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BrdU Labeling of Zebrafish Embryos

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Zebrafish embryos were incubated in E3 medium containing 10 mM of bromodeoxyuridine (BrdU) for 6 h, and then embryos were fixed with 4% PFA for 4 h, followed by 0.2 N of HCl for 1 h at room temperature. Anti-BrdU antibody (1:100; Santa Cruz Biotechnology, Paso Robles, CA, United States) and DAPI (1:100) were incubated overnight at 4°C. Embryos were washed with PBS and incubated with Goat Anti-Mouse IgG488 (1:200; Yisheng, Shanghai, China) overnight at 4°C. Histochemical detection was performed by Olympus MVX10 microscope (Olympus, Tokyo, Japan).

Chen S., Jia Z., Cai M., Ye M., Wu D., Wan T., Zhang B., Wu P., Xu Y., Guo Y., Tian C., Ma D, & Ma J. (2021). SP1-Mediated Upregulation of Long Noncoding RNA ZFAS1 Involved in Non-syndromic Cleft Lip and Palate via Inactivating WNT/β-Catenin Signaling Pathway. Frontiers in Cell and Developmental Biology, 9, 662780.

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