Overview

  • Product nameAnti-Ki67 antibody [EPR3610]
    See all Ki67 primary antibodies
  • Description
    Rabbit monoclonal [EPR3610] to Ki67
  • Tested applicationsIHC-Fr, WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Ki67 aa 1050-1150.

  • Positive control
    • WB: HeLa and ramos cell lysates. IHC-P: Human tonsil, colon, ovarian carcinoma, squamous cell carcinoma of cervix and colonic adenocarcinoma tissues. ICC/IF: HeLa, HT-29 cells, HAP1 cells. Flow Cyt: Ramos cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    This product is not suitable for xenograft experiments. For further information please contact our Customer Services team.

    Alternative versions available:

    Anti-Ki67 antibody (Alexa Fluor® 488) [EPR3610] (ab197234)
    Anti-Ki67 antibody (Alexa Fluor® 647) [EPR3610] (ab196907)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab92742 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-Fr Use at an assay dependent concentration.
WB 1/5000. Predicted molecular weight: 359 kDa.

For unpurified use at 1/500 - 1/1000.

IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/500 - 1/1000.

Flow Cyt 1/100 - 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionThought to be required for maintaining cell proliferation.
  • Sequence similaritiesContains 1 FHA domain.
  • Developmental stageExpression of this antigen occurs preferentially during late G1, S, G2 and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected.
  • Cellular localizationNucleus. Nucleus, nucleolus. Chromosome. Predominantly localized in the G1 phase in the perinucleolar region, in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix. In mitosis, it is present on all chromosomes.
  • Information by UniProt
  • Database links
  • Alternative names
    • Antigen identified by monoclonal antibody Ki 67 antibody
    • Antigen KI-67 antibody
    • Antigen KI67 antibody
    • KI67_HUMAN antibody
    • KIA antibody
    • Marker of proliferation Ki-67 antibody
    • MIB 1 antibody
    • MIB antibody
    • MKI67 antibody
    • PPP1R105 antibody
    • Proliferation related Ki 67 antigen antibody
    • Protein phosphatase 1 regulatory subunit 105 antibody
    • RP11-380J17.2 antibody
    see all

Anti-Ki67 antibody [EPR3610] images

  • ab92742 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92742 at 1µg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  • Anti-Ki67 antibody [EPR3610] (ab92742) at 1/5000 dilution (purified) + Ramos cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size : 359 kDa
    Observed band size : 395 kDa (why is the actual band size different from the predicted?)

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human squamous cell carcinoma of cervix tissue labelling Ki67 with purified ab92742 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Ki67 with purified ab92742 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Flow Cytometry analysis of Ramos cells labelling Ki67 with purified ab92742 at 1/150 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Overlay histogram showing Ramos cells stained with unpurified ab92742 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab92742, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic adenocarcinoma tissue labelling Ki67 with unpurified ab92742.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human normal colon tissue labelling Ki67 with unpurified ab92742.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling Ki67 with unpurified ab92742.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling Ki67 with unpurified ab92742.

  • Unpurified ab92742 staining Ki67 in human tonsil tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized and blocked with 4% serum + 1% BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/500 in blocking buffer) for 18 hours at 4°C. An Alexa Fluor® 568-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Magnification: 20X. Counterstained with DAPI.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling Ki67 with unpurified ab92742 at a dilution of 1/250.

  • Anti-Ki67 antibody [EPR3610] (ab92742) at 1/500 dilution (unpurified) + HeLa cell lysate at 10 µg

    Secondary
    HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 359 kDa
    Observed band size : 395 kDa (why is the actual band size different from the predicted?)

References for Anti-Ki67 antibody [EPR3610] (ab92742)

This product has been referenced in:
  • Ayuso JM  et al. Study of the Chemotactic Response of Multicellular Spheroids in a Microfluidic Device. PLoS One 10:e0139515 (2015). IF ; Human . Read more (PubMed: 26444904) »
  • Li J  et al. MiR-138 inhibits cell proliferation and reverses epithelial-mesenchymal transition in non-small cell lung cancer cells by targeting GIT1 and SEMA4C. J Cell Mol Med 19:2793-2805 (2015). IF ; Human . Read more (PubMed: 26283050) »

See all 9 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Marmoset (common) Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Colon
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Sep 25 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Colon
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Aug 19 2015

Application Flow Cytometry
Sample Human Cell (Differentiated hNSCs)
Permeabilization Yes - 0.25% Triton X-100 in DPBS
Gating Strategy Undifferentiated Stem Cells (white)
Specification Differentiated hNSCs
Preparation Cell harvesting/tissue preparation method: Accutase
Sample buffer: PBS
Fixation Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 15 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Tonsil)
Antigen retrieval step Heat mediated
Permeabilization No
Specification Tonsil
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Jun 18 2015

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Human Glioma into mouse brain)
Permeabilization Yes - Triton X-100
Specification Human Glioma into mouse brain
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted May 27 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step (agent) for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Human Cell (Breast cancer)
Specification Breast cancer
Permeabilization Yes - Triton X-100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 09 2014

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 22°C
Sample Human Tissue sections (Tonsil)
Specification Tonsil
Permeabilization Yes - 0.5%
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 28 2014

I tried to find the mouse and human specific skin cell markers, regrettably I couldn't find enough information about protein that are specific to human or mouse cells. I am sure further publication search will be helpful in answering this question. I w...

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ab92742 has indeed been tested in western blot on mouse and rat samples and gave a negative result.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"