Anti-BrdU antibody [BU1/75 (ICR1)] (ab6326)

Overview

• Product nameAnti-BrdU antibody [BU1/75 (ICR1)]See all BrdU primary antibodies ...
• Description
  Rat monoclonal [BU1/75 (ICR1)] to BrdU
• SpecificityThis antibody 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 with chlorodeoxyuridine but with reduced staining. The antibody does not react with thymidine. The antibody does not cross react with IdU.
• Tested applicationsICC/IF, IHC-FoFr, IHC-P, IHC (PFA fixed), ICC, IHC-Fr, Flow Cyt, IHC-FrFl more details
• Species reactivity
  Not applicable.
• Immunogen

  The details of the immunogen for this antibody are not available.

• General notesThe antibody recognises single stranded DNA so the DNA needs to be unraveled first. This can be done with DNAse, although this doesn't give the best results. Depending on the assay, acid denaturation with 2M HCL or heat denaturation are the most successful. Please note this step is critical in any assay with this antibody and is the area that should be modified to optimise results. Detailed BrdU protocol is available in "Neuroscience protocols" on our "Protocol and troubleshooting tips" webpage (www.abcam.com/protocols).

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.01% Sodium Azide
  Constituents: 50% Glycerol, PBS
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone numberBU1/75 (ICR1)
• IsotypeIgG2a
• Research Areas
  • Tags & Cell Markers
  • Cell Type Markers
  • Replication

  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • DNA / Nucleotides

  • Neuroscience
  • Cell Type Marker
  • Neuron marker
  • Soma marker

  • Cell Biology
  • Cell Cycle
  • Markers

Applications

Target

• RelevanceThe immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of tumor cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies is an accurate and comprehensive method to quantitate the degree of DNA-synthesis. BrdU is incorporated into the newly synthezised DNA of S-phase cells may provide an estimate for the fraction of cells in S-phase. Also dynamic proliferative information such as the S-phase transit rate and the potential doubling time can be obtained, by means of bivariate BrdU/DNA flow cytometric analysis.
• Cellular localizationNuclear

• Alternative names
  Bromodeoxyuridine antibodyBUdr antibody

Anti-BrdU antibody [BU1/75 (ICR1)] images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)
  Paraffin-embedded sections of hippocampus dentate gyrus stained for  BrdU-positive cells. The method is an indirect immunohistochemical technique: the primary antibody is ab6326 diluted 1/100 in PBS and the secondary antibody is an Alexa 488-conjugated anti-rat made in goat diluted 1/200.
  
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  Immunohistochemistry (Frozen sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)

  ab6326 used in double labeling on mouse sciatic nerve tissue (5µm) sections. This picture was kindly submitted by Dr Ulrich Hengst as part of his review on this product.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)This image is courtesy of an Abreview submitted by Dr Bing Lang
  ab6326 at 1/200 staining mouse E14.5 embryonic telencephalon tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retireval step was performed. The tissue was then stained with the antibody for 12 hours. A Texas Red conjugated donkey anti-rat antibody was used as the secondary.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)This image is courtesy of an anonymous Abreview

  ab6326 staining mouse adult epidermis tissue sections by IHC-P.  Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer prior to blocking in 10% serum for 1 hr at RT.  The primary antibody was diluted 1/200 and incubated with the sample for 24 hrs at 4°C.  A biotinylated goat anti-rat antibody was used as the secondary followed by a streptavidin-Cy3®.

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  Immunocytochemistry - BrdU antibody [BU1/75 (ICR1)] (ab6326)This image is courtesy of an anonymous Abreview

  ab6326 staining cultured cells of rat brain tissue by ICC.  The sample was PFA fixed and permeabilized in 1M HCl prior to blocking with 5% serum for 1 hour at 25°C.  The primary antibody was diluted 1/500 and incubated with the sample for 16 hours at 25°C.  A biotinylated rabbit anti-rat IgG antibody, diluted 1/200, was used as the secondary.

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  Immunohistochemistry (Frozen sections) - BrdU antibody [BU1/75 (ICR1)] (ab6326)This image is courtesy of an anonymous Abreview
  ab6326 staining BrdU in mouse brain tissue sections by IHC-Fr (paraformaldehyde-fixed frozen sections). Tissue samples were fixed with paraformaldehyde; permeabilized win 0.3% Triton X-100 and blocked with 5% Serum for 2 hours at 4°C. Before permeabilization samples were pretreated with 2N HCl at RT for 30 min and washed 3 times. The sample was incubated with primary antibody (1/200) at 4°C for 12 hours. An Alexa Fluor^® 488-conjugated Goat polyclonal to rat IgG (1/500) was used as secondary antibody. BrdU staining shown in green and NewN staining showin in red.

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• 

  Flow Cytometry - Anti-BrdU antibody [BU1/75 (ICR1)] (ab6326)This image is courtesy of an anonymous Abreview
  ab6326 staining BrdU in HeLa cells by Flow Cytometry. Cells were incubated with 10 µM BrdU for 30 minutes prior to being harvested with 1X trypsin-EDTA, washed twice in PBS containing 1% BSA, and fixed in 70% ethanol (added drop-wise) for at least 30 minutes on ice. Once fixed, pellets were acid denatured with HCl/Triton X-100 for 30 minutes at room temperature and then neutralised with sodium tetraborate.
  Pelleted cells were re-suspended in Tween/BSA/PBS to which primary antibody was then added (0.1 µg in 0.5% Tween 20 (v/v) plus 1% BSA in PBSA) and incubated for 30 minutes at room temperature. Secondary Alexa Fluor®488-conjugated Goat anti-Rat IgG (H+L) was used at 1/500 and incubated for 30 minutes at room temperature in the dark. Cells were pelleted once more and resuspended in PBS containing 5 µg/mL propidium iodide.
  Gating Strategy: Based on forward and side scatter, cells were gated into the region used for analysis. This was done by applying a large circle to accommodate the normal cells in an untreated control.

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