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BrdU antibody (Bromodeoxyuridine)

Details for Product anti-BrDU Antibody No. ABIN152983, Supplier: Log in to see
Antigen
Reactivity
Chemical
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Host
Rat
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Clonality (Clone)
Monoclonal ()
Conjugate
This BrdU antibody is un-conjugated
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Application
Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
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Options
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Immunogen Made to Chemical BrdU
Clone BU1-75 (ICR1)
Isotype IgG2a
Specificity This reacts with BrdU in single stranded DNA, BrdU attached to a protein carrier or free BrdU. It detects nucleated cells in S-Phase which have had BrdU incorporated into their DNA. Also reacts weakly with chlorodeoxyuridine, but does not cross react with thymidine or iododeoxyuridine.
Purification Protein G purified
Alternative Name Bromodeoxyuridine/BrdU
Target Type Chemical
Application Notes Western Blot, Flow Cytometry 1:25-1:200, Immunohistochemistry 1:10-1:500, Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry-Paraffin 1:25-1:200, Immunohistochemistry-Frozen 1:10-1:500Clone BU1/75 (ICR1) can be used for labeling paraffin-embedded tissue sections fixed in formalin. Denaturation of the DNA is critical for successful staining of BrdU. This can be achieved by exposing cells to heat, or acid. For heat-induced epitope retrieval, 10 mM citrate buffer pH 6.0 is recommended. Alternatively, a 30 min incubation in 2M HCl can be performed. The HCl must then be neutralized for 2 min with 0.1 M Na2B4O7. Pretreatment of tissues with proteinase K should be avoided. Immunohistochemistry-Frozen was reported in scientific literature. Use in Western blot reported in scientific literature (PMID 27425845).
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Protocol specific for BrdU Antibody Flow Cytometry Analysis: Prepare the following solutions before proceeding:Phosphate buffered saline (PBS)2N HCl, 0.5 % Triton X-100PBS containing 0.05 % Tween-20PBS containing 1 % BSA (PBS/BSA)10mg/ml Propidium iodide (PI)As a positive control, BrdU labeled cells maybe obtained from Phoenix Flow Systems (http://www.phnxflow.com), catalogue number ACNC12.
1. Add BrdU to the cell suspension in culture medium to a final concentration of 10 umol/L and incubate for 30 minutes in a CO2 incubator at 37C.
. Wash cells twice with PBS/BSA, and resuspend in PBS
. Add cells slowly into 5ml of 70 % ethanol at -20C, mixing continuously (vortex preferred). Incubate on ice for 30 minutes.
. Centrifuge at 500g for 10 minutes, decant supernatant, and resuspend cell pellet.
. Add 2ml 2N HCl, 0.5 % Triton X-100 and incubate the cells for 30 minutes at room temperature (preferably on a rocking platform).
. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend in 3 mL 0.1M Na2B4O7, pH 8.
. Centrifuge at 500g for 10 minutes, decant supernatant and resuspend the cells in PBS/BSA + 0.05 % Tween
. Adjust cell concentration to 1 x 10(7)/ml
. Aliquot 100ul of cell suspension into required number of 12 x 75mm tubes.
. Incubate the cells with the anti-BrdU antibody at the recommended dilution for 30 minutes at room temperature.
. Add 2 mLs PBS/BSA and centrifuge the cells at 1000rpm for 5 minutes.
. If a secondary antibody layer is required then decant the wash and incubate the cells with the secondary antibody for 30 minutes at room temperature. If no secondary antibody layer is required then proceed to step 13.
. Wash the cells after the secondary antibody layer by repeating step 10.
. Decant off the wash and add 1ml PBS containing 10ug/ml PI (Dilute the 10mg/ml solution of PI 1/1000 in a suitable volume of PBS)
. Analyse cells by flow cytometry following the manufacturer's instructions. The PI should be read on the appropriate channel set to the Peak/Area and not log scale.IMMUNOHISTOLOGY Formalin-fixed paraffin-embedded tissue sections:Clone BU1/75 (ICR1) can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pretreatment of tissues with heat-induced epitope retrieval using 10mM citrate buffer pH6.0 is recommended. Pretreatment of tissues with proteinase K should be avoided.
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS
Buffer contains: 0.09 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C,-20 °C
Storage Comment Store at 4°C short term. Aliquot and store at -20°C long term. Avoid freeze-thaw cycles.
Supplier Images
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN1...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN1...
Flow Cytometry (FACS) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN152983] - HL-60 ...
Flow Cytometry (FACS) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Flow Cytometry: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN152983] - Analys...
Immunofluorescence (IF) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Immunocytochemistry/Immunofluorescence: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN1...
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) image for anti-BrdU antibody (Bromodeoxyuridine) (ABIN152983) Immunohistochemistry-Paraffin: Bromodeoxyuridine/BrdU Antibody (BU1/75 (ICR1)) [ABIN1...
Product cited in: Gygli, Chang, Gokozan, Catacutan, Schmidt, Kaya, Goksel, Baig, Chen, Griveau, Michowski, Wong, Palanichamy, Sicinski, Nelson, Czeisler, Otero: "Cyclin A2 promotes DNA repair in the brain during both development and aging." in: Aging, Vol. 8, Issue 7, pp. 1540-70, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Western Blotting,Immunohistochemistry

Ishikawa, Fukushima, Frankland, Kida: "Hippocampal neurogenesis enhancers promote forgetting of remote fear memory after hippocampal reactivation by retrieval." in: eLife, Vol. 5, 2016 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry

Seidel, Remus, Delacher, Grigaravicius, Reuss, Frappart, von Deimling, Feuerer, Abdollahi, Frappart: "Epidermal Nbn deletion causes premature hair loss and a phenotype resembling psoriasiform dermatitis." in: Oncotarget, Vol. 7, Issue 17, pp. 23006-18, 2016 (PubMed). Method employed by authors: Western Blotting (WB)

Lamm, Maoz, Bester, Im, Shewach, Karni, Kerem: "Folate levels modulate oncogene-induced replication stress and tumorigenicity." in: EMBO molecular medicine, Vol. 7, Issue 9, pp. 1138-52, 2015 (PubMed). (Sample species: Human).

Somyajit, Saxena, Babu, Mishra, Nagaraju: "Mammalian RAD51 paralogs protect nascent DNA at stalled forks and mediate replication restart." in: Nucleic acids research, Vol. 43, Issue 20, pp. 9835-55, 2015 (PubMed). (Sample species: Human). Further details: Immunocytochemistry,Immunofluorescence

Feng, Liang, Li, Li, Liang, Zhao, McNutt, Yin: "PTEN Controls the DNA Replication Process through MCM2 in Response to Replicative Stress." in: Cell reports, Vol. 13, Issue 7, pp. 1295-303, 2015 (PubMed).

Zhu, Su, Mukherjee, Mori, Hu, Asaithamby: "FANCD2 influences replication fork processes and genome stability in response to clustered DSBs." in: Cell cycle (Georgetown, Tex.), Vol. 14, Issue 12, pp. 1809-22, 2015 (PubMed).

Harmacek, Watkins-Chow, Chen, Jones, Pavan, Salbaum, Niswander: "A unique missense allele of BAF155, a core BAF chromatin remodeling complex protein, causes neural tube closure defects in mice." in: Developmental neurobiology, Vol. 74, Issue 5, pp. 483-97, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunocytochemistry,Immunofluorescence,Immunohistochemistry

Xie, Yen, Owonikoko, Ramalingam, Khuri, Curran, Doetsch, Deng: "Bcl2 induces DNA replication stress by inhibiting ribonucleotide reductase." in: Cancer research, Vol. 74, Issue 1, pp. 212-23, 2014 (PubMed). Further details: Immunocytochemistry,Immunofluorescence

Di Giovannantonio, Di Salvio, Omodei, Prakash, Wurst, Pierani, Acampora, Simeone: "Otx2 cell-autonomously determines dorsal mesencephalon versus cerebellum fate independently of isthmic organizing activity." in: Development (Cambridge, England), Vol. 141, Issue 2, pp. 377-88, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Toko, Hariharan, Konstandin, Ormachea, McGregor, Gude, Sundararaman, Joyo, Joyo, Collins, Din, Mohsin, Uchida, Sussman: "Differential regulation of cellular senescence and differentiation by prolyl isomerase Pin1 in cardiac progenitor cells." in: The Journal of biological chemistry, Vol. 289, Issue 9, pp. 5348-56, 2014 (PubMed). Further details: Flow Cytometry

Conway, Schaffer: "Biomaterial microenvironments to support the generation of new neurons in the adult brain." in: Stem cells (Dayton, Ohio), Vol. 32, Issue 5, pp. 1220-9, 2014 (PubMed).

Cai, Sugimoto, Arainga, Alvarez, Didier, Kuroda: "In vivo characterization of alveolar and interstitial lung macrophages in rhesus macaques: implications for understanding lung disease in humans." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 192, Issue 6, pp. 2821-9, 2014 (PubMed). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Nobs, Baranek, Nestel, Kulik, Kapfhammer, Nitsch, Atanasoski: "Stage-specific requirement for cyclin D1 in glial progenitor cells of the cerebral cortex." in: Glia, Vol. 62, Issue 5, pp. 829-39, 2014 (PubMed). Further details: Immunohistochemistry (Frozen Sections)

Ishikawa, Kim, Namba, Kohsaka, Uchino, Kida: "Time-dependent enhancement of hippocampus-dependent memory after treatment with memantine: Implications for enhanced hippocampal adult neurogenesis." in: Hippocampus, Vol. 24, Issue 7, pp. 784-93, 2014 (PubMed). Further details: Immunohistochemistry (Frozen Sections)

Kawaue, Sagou, Kiyonari, Ota, Okamoto, Shinoda, Kawaguchi, Miyata: "Neurogenin2-d4Venus and Gadd45g-d4Venus transgenic mice: visualizing mitotic and migratory behaviors of cells committed to the neuronal lineage in the developing mammalian brain." in: Development, growth & differentiation, Vol. 56, Issue 4, pp. 293-304, 2014 (PubMed). Further details: Immunohistochemistry (Frozen Sections)

Diensthuber, Zecha, Wagenblast, Arnhold, Edge, Stöver: "Spiral ganglion stem cells can be propagated and differentiated into neurons and glia." in: BioResearch open access, Vol. 3, Issue 3, pp. 88-97, 2014 (PubMed).

Quadrato, Elnaggar, Duman, Sabino, Forsberg, Di Giovanni: "Modulation of GABAA receptor signaling increases neurogenesis and suppresses anxiety through NFATc4." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 34, Issue 25, pp. 8630-45, 2014 (PubMed). Further details: Immunohistochemistry (Free Floating)

Tong, Han, Shah, Obernier, Guinto, Alvarez-Buylla: "Primary cilia are required in a unique subpopulation of neural progenitors." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, Issue 34, pp. 12438-43, 2014 (PubMed). (Sample species: Mouse (Murine)). Further details: Immunohistochemistry (Paraffin-embedded Sections)

Lee, Kim, Chao, Kuo, Mally, McClean, Pemberton, Wilmington, Wong, Murphy: "Progesterone and allopregnanolone improve stroke outcome in male mice via distinct mechanisms but neither promotes neurogenesis." in: Journal of neurochemistry, 2014 (PubMed). Further details: Immunohistochemistry (Frozen Sections)