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  1. Link

    brdu-antibody-iib5-ab8152.pdf

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Anti-BrdU antibody [IIB5] (ab8152)

  • Datasheet
  • SDS
Reviews (7)Q&A (5)References (57)

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Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)
  • Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)

Key features and details

  • Mouse monoclonal [IIB5] to BrdU
  • Suitable for: IHC-FoFr, ICC/IF, Flow Cyt, IHC-Fr, IHC-P
  • Reacts with: Species independent
  • Isotype: IgG1

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Biochemical
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BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (ab142567)
Primary
Product image
Alexa Fluor® 647 Anti-BrdU antibody [BU1/75 (ICR1)] (ab220075)

View more associated products

Overview

  • Product name

    Anti-BrdU antibody [IIB5]
    See all BrdU primary antibodies
  • Description

    Mouse monoclonal [IIB5] to BrdU
  • Host species

    Mouse
  • Specificity

    BrdU is a thymidine analogue and when offered to proliferating cells it is incroporated into reduplicating cells. The antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.
  • Tested applications

    Suitable for: IHC-FoFr, ICC/IF, Flow Cyt, IHC-Fr, IHC-Pmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule corresponding to BrdU conjugated to bovine serum albumin.

  • Positive control

    • Bromodeoxyuridine labeled cells.
  • General notes

    The following product is available as purified antibody (purified by affinity chromatography) together with several conjugated versions:

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

    Unstained positive control slides from mice treated with BrdU (formalin-fixed, paraffin-embedded intestine sections) are available as BrdU control slides ab129956.

     

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    pH: 7.3
    Preservative: 0.09% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    IIB5
  • Myeloma

    Sp2/0-Ag14
  • Isotype

    IgG1
  • 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

  • Alternative Versions

    • Anti-BrdU antibody [IIB5] (ab8955)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Related Buffer

    • BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (ab142567)
  • Related Products

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8152 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-FoFr (1)
Use at an assay dependent concentration.
ICC/IF (4)
Use at an assay dependent concentration.
Flow Cyt
1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr
1/5 - 1/10.

Dilute antibody in 0.15 M phosphate buffered saline with 1% BSA and 1% Na-azide.

IHC-P (2)
1/5 - 1/10. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Enzymatic antigen retrieval with proteases can also be used.

Notes
IHC-FoFr
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr
1/5 - 1/10.

Dilute antibody in 0.15 M phosphate buffered saline with 1% BSA and 1% Na-azide.

IHC-P
1/5 - 1/10. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Enzymatic antigen retrieval with proteases can also be used.

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

  • Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)
    Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
    ab8152 (1/100) staining BrdU in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). For further esperimental details please see Abreview.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)This image is courtesy of an anonymous Abreview
    Immunohistochemical analysis of rat brain tissue, staining BrdU with ab8152.

    Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/20 in diluent) for 18 hours at 25°C. A Cy3®-conjugated donkey anti-mouse polyclonal IgG was used as the secondary antibody.

    See Abreview

  • Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)
    Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)

    Cells were pulse labeled with 10 µM BrdU for 30 min, rinsed twice in prewarmed PBS, and chased in prewarmed culture medium, supplemented with 5 mM deoxythymidine. Incorporated BrdU was detected after ethanol fixation of the cells, which were than rinsed once in PBS and resuspended in 2 ml of 0.4 mg/ml pepsin in 0.1 N HCl. After 30 min at room temperature cells were pelleted, resuspended in 2 N HCl, and incubated for another 30 min at 37°C. Cells were rinsed in 0.1 M borate buffer, pH 8.5, and PBS/BSA (1 mg/ml BSA in PBS). Appropriately diluted mouse anti-BrdU antibody (clone IIB5) was added to the cell pellet, resuspended in 100 micro liters PBS/BSA. After incubation for 1 h at room temperature, the cells were rinsed twice in PBS/BSA. For visualization, FITC-conjugated Fab2 fragments of rabbit anti-mouse Ig antibody were added in a 1/10 dilution. After incubation for 45 min at room temperature samples were rinsed twice in PBS/BSA and the cells were finally resuspended in 0.5 ml cold PBS supplemented with 100 microgram/ml RNAse and 20 µg/mL propidium iodide. The samples were allowed to stand for 15 min on ice in the dark before flow cytometric analysis. In the negative control the primary antibody was omitted.

Protocols

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (57)

Publishing research using ab8152? Please let us know so that we can cite the reference in this datasheet.

ab8152 has been referenced in 57 publications.

  • Zhou D  et al. Acarbose ameliorates spontaneous type-2 diabetes in db/db mice by inhibiting PDX-1 methylation. Mol Med Rep 23:N/A (2021). PubMed: 33236139
  • Feng X  et al. Transplanted embryonic retinal stem cells have the potential to repair the injured retina in mice. BMC Ophthalmol 21:26 (2021). PubMed: 33422026
  • Yang G & Zhao Y MicroRNA-490-3p inhibits inflammatory responses in LPS-induced acute lung injury of neonatal rats by suppressing the IRAK1/TRAF6 pathway. Exp Ther Med 21:152 (2021). PubMed: 33456519
  • Aichen Z  et al. LncRNA FGD5-AS1 promotes the malignant phenotypes of ovarian cancer cells via targeting miR-142-5p. Apoptosis 26:348-360 (2021). PubMed: 33974163
  • Pavel M  et al. a-Catenin levels determine direction of YAP/TAZ response to autophagy perturbation. Nat Commun 12:1703 (2021). PubMed: 33731717
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 12 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody [IIB5]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Intestine)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 2M HCl
Permeabilization
Yes - Triton X-100
Specification
Intestine
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde
Read More

DR. Egi Kardia

Verified customer

Submitted Sep 21 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU antibody [IIB5]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (Primary airway epithelial cells)
Permeabilization
No
Specification
Primary airway epithelial cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
70% Ethanol
Read More

DR. Egi Kardia

Verified customer

Submitted Sep 06 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU antibody [IIB5]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Sample
Human Cell (OCI ly1)
Specification
OCI ly1
Permeabilization
Yes - 0.25% Triton X-100 in PBS
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 07 2014

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-BrdU [IIB5] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (brain)
Specification
brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Dec 07 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU [IIB5] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
African Green Monkey Cell (Oligodendrocytes, astrocytes (mixed glial culture))
Specification
Oligodendrocytes, astrocytes (mixed glial culture)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton-PBS for 30 minutes followed by DNAse treatment for 90 minutes.
Read More

Abcam user community

Verified customer

Submitted Oct 28 2010

Question

Hello, I need to work with IdU and CldU for double staining. I found out that I can use for cldU the ab6326 that doen't cross react with IdU. Can I use ab8039 or ab8152 for IdU? I need an antibody that doesn't cross react with CldU.

Read More

Abcam community

Verified customer

Asked on Apr 03 2013

Answer

Thank you for your inquiry and for rating your experience with us.

I just wanted to follow up on your query with some additional information.

For ab8039, this antibody can be used for detection of BrdU and BrU. We have not tested it regarding other chemicals of this family.

For ab8152, this antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.

I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Apr 03 2013

Question

the floating sections are not frozen.all steps are performed with the sections positioned in12 well plates.

it is not possible to work withthem on slides because they immediately come off . once they are immersed in citric buffer they will surely come off.

i ordered the HIER solution you offered , but i wonder how to work with it in the conditions specified above.

Read More

Abcam community

Verified customer

Asked on Jun 19 2012

Answer

Thank you for contacting us.

For floating sections, please check thefollowing protocol on IHC World website:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm
This protocol is for paraffin-embedded as well as frozen sections.

As for ensuring the paraffin sections stick to your slides:
Different sections and tissues stick with differetn strength to the slides.
It might be advisable to use charged slides, as well as to bake your sections onto the slides by exposing them to heat (e.g. heating plate or oven) up to 60 degree Celsius.The purpose is to remove miscroscopic amounts of water between the sections and the slide.
Subsequently the paraffin would need to removed according to a standard IHC-P protocol. Then you would proceed with the antigen retrieval step.

Also, please check this information on the IHC World website for further advice.
http://www.ihcworld.com/_faq/histology-faq/section/s1.htm

Read More

Abcam Scientific Support

Answered on Jun 19 2012

Question

how do i technically imerse the tissue sedctions in the citrate buffer without loosing them?

if sections are on a slide, and the slide is imersed in the boiling buffer, the sections will detach from the slides.

also, i am working with floating sections. do ypu have a technical tip re howthis step should be excecuted?

Read More

Abcam community

Verified customer

Asked on Jun 14 2012

Answer

The slides need to be immersed, for the retrieval to be effective. Yes, the sections could come off ifthey are kept too long in the retrieval solution. Should they come off, you would need to reduce the time they are exposed to the retrieval buffer.

Are the floating sections paraffin-embedded or frozen? I have found the following protocol on IHC World website which should help you further:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm

Read More

Abcam Scientific Support

Answered on Jun 14 2012

Question

Inquiry: what type of antigen retreival is required in order to detect the brdu in PFA sixed tissue sections, along with doublecortin detection? the protocol that uses HCl does not allow dapi counterstain.and also destroys the other antigen ,which is cytosolic. if citric acid should be used- can you send me the excat protocol? including steps of trypsin, etc. which secondary Ab should be used and at what concentration?

Read More

Abcam community

Verified customer

Asked on Jun 05 2012

Answer

I have checked with the lab:
Antigen retrieval can be done with HIER using citrate buffer.

Please see below for the protocol:

High Temperature antigen Unmasking Technique using Sodium Citrate Buffer for Immunohistochemical Demonstration on Paraffin Sections

1. Cut and mount sections on slides coated with Vectabond or APES (3-aminopropyltriethoxysilane).
2. Deparaffinize sections and rehydrate to distilled water.
3. Bring 1600ml 0.01M sodium citrate buffer (pH 6.0) to the boil in a Prestige stainless steel pressure cooker, using a hot plate. Cover but do not lock lid.
4. Position slides into metal staining racks and lower into pressure cooker ensuring slides are well immersed in citrate buffer. Lock lid. The small valve will rise.
5. When the pressure indicator valve (the large one) has risen after about 4 minutes, incubate sections for 1 minute.
6. Remove pressure cooker from heat source and run under cold water with lid on. When the small valve sinks open lid and remove slides and place immediately into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE SINKS.
7. Wash sections in TBS buffer (pH 7.6) for 1 x 5 minutes.
8. Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes.
9. Wash sections in distilled water for 2 x 5 minutes, then wash sections in TBS buffer for 2 x 5 minutes.
10. Place sections in normal serum for 20 minutes.
11. Cover sections with primary antibody. ( The optimal dilution of the antibody, incubation time and incubation temperature should be determined by the individual laboratory).
12. Wash in TBS buffer for 2 x 5 minutes.
13. Incubate sections in secondary antibody for 30 minutes.
14. Wash in TBS buffer for 2 x 5 minutes.
15. Incubate slides in ABComplex for 30 minutes.
16. Wash in TBS buffer for 2 x 5 minutes.
17. Incubate slides in DAB.
18. Wash in water for 2 x 5 minutes.
19. Counterstain with haematoxylin (if required), dehydrate, coverslip and mount.


To avoid sections becoming detached, sections should be mounted on Vectabond or APES covered slides, then dried at 37oC overnight followed by drying at 56oC for 60 minutes.

Read More

Abcam Scientific Support

Answered on Jun 05 2012

Question

Do you know how long should be the ssDNA to be detected with the ab by Immunofluorescence? I want to use is to detect fragment between 30-300 nt fron adherent cells in culture.

Read More

Abcam community

Verified customer

Asked on Nov 18 2011

Answer

Thank you for contacting us with your question. We unfortunately do not have any data regarding the minimum length of ssDNA required for antibody staining on adherent cells.

Read More

Abcam Scientific Support

Answered on Nov 18 2011

1-10 of 12 Abreviews or Q&A

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