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  1. Link

    biotin-brdu-antibody-ab2284.pdf

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Biotin Anti-BrdU antibody (ab2284)

  • Datasheet
Reviews (6)Q&A (7)References (42)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)
  • Immunocytochemistry/ Immunofluorescence - Biotin Anti-BrdU antibody (ab2284)
  • Immunohistochemistry (Frozen sections) - Biotin Anti-BrdU antibody (ab2284)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)

Key features and details

  • Biotin Sheep polyclonal to BrdU
  • Suitable for: IHC-P, IHC-Fr, ICC/IF, ELISA, IHC-FoFr
  • Reacts with: Species independent
  • Conjugation: Biotin
  • Isotype: IgG

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BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
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Overview

  • Product name

    Biotin Anti-BrdU antibody
    See all BrdU primary antibodies
  • Description

    Biotin Sheep polyclonal to BrdU
  • Host species

    Sheep
  • Conjugation

    Biotin
  • Tested applications

    Suitable for: IHC-P, IHC-Fr, ICC/IF, ELISA, IHC-FoFrmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Bromodeoxyuridine coupled to keyhole limpet hemocyanin (KLH)

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.50
    Constituent: 0.4% PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Markers
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Replication
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides

Associated products

  • Related Buffer

    • BrdU Cell Proliferation ELISA Kit (chemiluminescent) (ab126572)
  • Related Products

    • BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    • BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (ab142567)
    • Biotin Assay Kit (Colorimetric) (ab185441)
    • Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)

Applications

Our Abpromise guarantee covers the use of ab2284 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. Fixation in cold methanol for 30 minutes followed by immersion in 7 x 10-3 N NaOH for 10-15 seconds allows BrdU staining with the simultaneous detection of nuclear cytoplasmic and membrane assigns as well as preservation of morphological detail.
ICC/IF Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration. dilute from 1/500 to 1/40,000 against 1mg/mL BrdU analyte.
IHC-FoFr 1/2000. 1/2000 (see Abreview).

Target

  • Relevance

    The 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 localization

    Nuclear
  • Alternative names

    • Bromodeoxyuridine antibody
    • BUdr antibody

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)This image is courtesy of an anonymous Abreview

    ab2284 staining BrdU in mouse intestine tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% H2O2 in methanol for 12 minutes; antigen retrieval was by heat mediation in 10mM citrate, pH6. Samples were incubated with primary antibody (1/100) for 24 hours at 4°C.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Biotin Anti-BrdU antibody (ab2284)
    Immunocytochemistry/ Immunofluorescence - Biotin Anti-BrdU antibody (ab2284)This image is courtesy of an anonymous Abreview

    ab2284 at 1/250 staining primary E12 mouse cortex cells by ICC/IF. The cells were paraformaldehyde fixed, blocked with serum and then incubated with the antibody for 24 hours. Streptavidin conjugated to Alexa-Fluor ®  488 was used as the secondary. The image shows BrdU staining with nuclei counterstained with DAPI.

    See Abreview

  • Immunohistochemistry (Frozen sections) - Biotin Anti-BrdU antibody (ab2284)
    Immunohistochemistry (Frozen sections) - Biotin Anti-BrdU antibody (ab2284)This image is courtesy of an Abreview submitted by Ahmar Aziz

    ab2284 staining BrdU in Mouse skin tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 30 minutes. Samples were incubated with primary antibody (1/50 in PBS) for 12 hours at 4°C. A Streptavidin Alexa Fluor® 488-conjugated Goat polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-BrdU antibody (ab2284)This image is courtesy of an Abreview submitted by Dr Christoph Schwarzer
    ab2284 at 1/2000 dilution staining mouse free floating brain slices by Immunohistochemistry (Formalin/PFA fixed sections). The mice were treated with 100mg/kg BrdU 2 hours before fixation. Free floating 40µm vibratome sections were obtained from paraformaldehyde fixed brains, these were incubated with the antibody for 24 hours. A streptavidin-HRP complex and DAB were used for detection. The image depicts the subventricular zone.

    See Abreview

Protocols

  • ab2284 IHC-P recommended protocol

Click here to view the general protocols

Datasheets and documents

    • Datasheet
  • References (42)

    Publishing research using ab2284? Please let us know so that we can cite the reference in this datasheet.

    ab2284 has been referenced in 42 publications.

    • Fu DJ  et al. Gastric squamous-columnar junction contains a large pool of cancer-prone immature osteopontin responsive Lgr5-CD44+ cells. Nat Commun 11:84 (2020). PubMed: 31901081
    • Xiao Y  et al. LncRNA MALAT1 increases the stemness of gastric cancer cells via enhancing SOX2 mRNA stability. FEBS Open Bio 9:1212-1222 (2019). PubMed: 31037832
    • Flentke GR  et al. Alcohol-mediated calcium signals dysregulate pro-survival Snai2/PUMA/Bcl2 networks to promote p53-mediated apoptosis in avian neural crest progenitors. Birth Defects Res N/A:N/A (2019). PubMed: 31021056
    • Razumilava N  et al. Hedgehog Signaling Modulates Interleukin-33-Dependent Extrahepatic Bile Duct Cell Proliferation in Mice. Hepatol Commun 3:277-292 (2019). PubMed: 30766964
    • Campanale JP  et al. Methods to label, isolate, and image sea urchin small micromeres, the primordial germ cells (PGCs). Methods Cell Biol 150:269-292 (2019). PubMed: 30777180
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 13 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-BrdU antibody (Biotin)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Skin)
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10%
    Fixative
    Acetone
    Read More

    Ahmar Aziz

    Verified customer

    Submitted Feb 10 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody (Biotin)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM citrate, pH6
    Sample
    Mouse Tissue sections (intestine)
    Specification
    intestine
    Blocking step
    (agent) for 12 minute(s) · Concentration: 1%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 09 2014

    Question

    May I put this antibody in -800C for storage?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 29 2012

    Answer

    This antibody can be at -80C in aliquots for storage, and this may extend the life of the antibody. Either -20C or -80C will be fine.

    Read More

    Abcam Scientific Support

    Answered on Mar 29 2012

    Question

    I use your protocol for BrdU staining. Please see attached file. I used your antibody, ab2284 on mice intestine tissue last November, I got beautiful staining. This month, we are doing another batch of mice and following exactly protocol as November’s. I did three run of staining to exclude any mistake I might have and still got no stain at all. Finally, I thought it was DAB problem and emailed you last week. After receiving new DAB, I repeat BrdU stain with a fresh aliquated antibody. Then I got faint stain - comparing with last November’s staining. At that time, I thought it was Streaptavidin problem and emailed you for that. Since I need result quickly, I run another run with my real slides on Monday with the antibody I thawed last Thursday (got result on Friday) and got no stain at all even with new DAB. This antibody was aliquated and stored at -200C since we received last September. This particular tube was thawed last Thursday and used only once. It was put back to -200C right after usage. This Monday was the second time to thaw and to use, and I did not get any staining, the only explanation is that the antibody got degraded.

    I hope you agree with me or give me some suggestion. I also hope you could send me new antibody to see what I get.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 29 2012

    Answer

    Thank you for your reply and forsending the additional information.

    It does sound like the antibody was damaged during storage, so I am happy to send a replacement vial for you. This will be coming free of charge on the order ***, which should arrive tomorrow. Please note that we recommend storing this antibody in working aliquots, so that each aliquot is not frozen and thawed more than one time.

    I apologize for the trouble with these products, but please keep me updated about how this replacement antibody works in your staining. Let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Mar 29 2012

    Question

    We want to use the biotinylated anti-BrdU antibody without a secondary antibody. I was wondering if you would suggest blocking the tissue with hydrogen peroxide or denaturing the DNA with HCl first?

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2012

    Answer

    You will denature the DNA first and then carry on with the IHC staining protocol.

    https://www.abcam.com/index.html?pageconfig=resource&rid=11533
    https://www.abcam.com/assets/popups/popup_datasheet_protocols_printer_friendly.cfm?pid=1325&intAbId=2284

    Read More

    Abcam Scientific Support

    Answered on Feb 22 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody (Biotin)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Mouse tumour)
    Specification
    Mouse tumour
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.1% Triton X-100
    Blocking step
    Perkin-Elmer TNB Blocking Reagent as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 22 2008

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody (Biotin)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (brain,lateral ventricle)
    Specification
    brain,lateral ventricle
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: Tris Buffered Saline- 0.3% Triton X
    Permeabilization
    Yes - 4N HCl
    Blocking step
    NDS as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Mr. stefano gaburro

    Verified customer

    Submitted Nov 01 2007

    Question

    Do you know if somebody try to precipitate chromatin with BrdU antibody?

    Read More

    Abcam community

    Verified customer

    Asked on Apr 18 2007

    Answer

    I managed to find some references that you might be interested in. 1) Hiraga,et al. 2005. DNA polymerases alpha, delta, and epsilon localize and function together at replication forks in Saccharomyces cerevisiae. Genes to Cells. 10, 297-309. (http://www.genestocellsonline.org/cgi/content/full/10/4/297) 2) Lin, et al. 2006. Inhibition of estradiol-induced mammary proliferation by dibenzoylmethane through the E2–ER–ERE-dependent pathway. Carcinogenesis. 27(1):131-136

    Read More

    Abcam Scientific Support

    Answered on Apr 18 2007

    Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU antibody (Biotin)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Primary E12 mouse cortex)
    Specification
    Primary E12 mouse cortex
    Fixative
    Paraformaldehyde
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 22 2007

    Question

    Is the borate buffer made with just boric acid or boric acid plus Nabicarbonate? I have found to borate buffer recipies see below. I did try looking at the ref on the website from you IHC borate wash buffer but it is not accessible online. I assume the buffer is just quenching the acid? Borate buffer 1. 1M Borate Buffer 100mls: 6.183g boric acid, 70mls dh2O and ph to pH8.0 with 10N NaOH Dilute 1:10 with dH2O to give 0.1M borate buffer. 2. Boric Acid-Borate Buffer (pH 7.6) Stock Solution A: Boric Acid 1.24 gm/100 ml distilled water Stock Solution B: Sodium Biborate 1.9 gm/100 ml distilled water Solution A ......... 85 ml Solution B ......... 15 ml Mix solutions A and B. Stable at room temperature. Supplies: Boric acid (H3BO3) Mallinckrodt Specialty Chemical Co. http://www.mallinckrodt.com/ Paris, KY Phone: 1-314-654-2000 Catalog No. 2549 Sodium biborate (Na2B4O7.10H2O) J.T. Baker Inc. http://www.jtbaker.com/ Phillipsburg, NJ Phone: 1-800-582-2537 Catalog No. 3570-01 Thankyou for your help Best wishes

    Read More

    Abcam community

    Verified customer

    Asked on Jan 16 2007

    Answer

    Thank you for your question. I'm afraid as I have not done the experiments I don't have a recipe for the borate buffer. I find that IHC world (www.IHCworld.com) has often some good protocols, and the following link (http://www.ihcworld.com/_protocols/antibody_protocols/brdu_accurate_chem_sci.htm) mentions 0.1M Borate Buffer, pH 8.5 made with Sodium borate (MW 381.4) 3.8 g + Distilled water 100 ml. Indeed the Borate buffer is simply to neutralize the acid.

    Read More

    Abcam Scientific Support

    Answered on Jan 16 2007

    1-10 of 13 Abreviews or Q&A

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