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Anti-Ki67 antibody [PP-67] - Proliferation Marker (ab6526)

Overview

  • Product nameAnti-Ki67 antibody [PP-67] - Proliferation MarkerSee all Ki67 primary antibodies ...
  • Description
    Mouse monoclonal [PP-67] to Ki67 - Proliferation Marker
  • SpecificityMonoclonal Anti-Proliferating Cell Protein Ki-67 reacts with the Ki-67 nuclear antigen (345 and 395 kD, and several lower molecular weight proteins) in immuno-blotting. The product shows an immunohistochemical staining pattern similar to that obtained with other anti-Ki-67/MIB-1 reagents. It labels proliferating cells in frozen human tissue and in formalin-fixed, paraffin-embedded tissue. A microwave heating retrieval step is required for successful labeling of routinely formalin-fixed, paraffin-embedded human tissue.
  • Tested applicationsICC/IF, IHC-P, WB, IHC-Fr, Flow Cyt more details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A HeLa cell nuclear preparation.

  • Positive controlHuman Tonsil

Properties

Applications

Our Abpromise guarantee covers the use of ab6526 in the following tested applications.

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

Application Notes
ICC/IF ICC/IF: 1/200. (see review). Fix with 4% PFA. Permeabilisation may be necessary (e.g triton, methanol).
IHC-P IHC-P: Use at an assay dependent dilution. We suggest an incubation period of 30 minutes at room temperature. High temperature treatment of formalin-fixed tissue sections with 10 mM citrate buffer, pH 6.0 must be performed prior to the immunostaining.
WB WB: Use at an assay dependent dilution.
IHC-Fr IHC-Fr: 1/800. (see review). Fix with formalin. Permeabilisation may be necessary (e.g triton, methanol).
Flow Cyt Flow Cyt: 1/100.

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.
  • Target information above from: UniProt accession P46013 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
      Antigen identified by monoclonal antibody Ki 67 antibodyAntigen KI-67 antibodyAntigen KI67 antibody
      KI67_HUMAN antibodyKIA antibodyMKI67 antibodyProliferation related Ki 67 antigen antibodyRP11-380J17.2 antibody
    see all

Anti-Ki67 antibody [PP-67] - Proliferation Marker images

  • ab6526 at a 1/500 dilution staining PBMCs from human tonsil tissue sections by IHC-P. The tissue was paraformaldehyde fixed and blocked with 10% goat serum and then incubated with the antibody overnight at 4°C. A biotinylated goat anti-mouse IgG (H&L) was used as the secondary antibody.

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  • ab6526 staining normal human primary fibroblast cells by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking with 10% BSA for 2 hours at 25°C.  The primary antibody was diluted 1/100 and incubated with the sample for 2 hours at 25°C.  A FITC conjugated goat anti-mouse IgM antibody was used as the secondary.

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  • Overlay histogram showing Ramos cells stained with ab6526 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab6526, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Ramos cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining Ki67 with ab6526.

References for Anti-Ki67 antibody [PP-67] - Proliferation Marker (ab6526)

This product has been referenced in:
  • Pruvot B  et al. Leukemic cell xenograft in zebrafish embryo for investigating drug efficacy. Haematologica 96:612-6 (2011). IHC-P . Read more (PubMed: 21228037) »
  • Ortega JA & Alcántara S BDNF/MAPK/ERK-induced BMP7 expression in the developing cerebral cortex induces premature radial glia differentiation and impairs neuronal migration. Cereb Cortex 20:2132-44 (2010). Read more (PubMed: 20038543) »

See all 8 Publications for this product

Product Wall

Displaying 1 - 10 of 11 results for Abreviews and Q&A

Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (brain (lateral ventricles), SVZ)
Specification brain (lateral ventricles), SVZ
Fixative Paraformaldehyde
Permeabilization Yes - triton 0.5%
Blocking step chemiblocker as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 8°C
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Verified customer

Submitted Nov 04 2009

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (brain (lateral ventricles))
Specification brain (lateral ventricles)
Fixative Paraformaldehyde
Permeabilization Yes - triton 5%
Blocking step chemiblocker as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: 8°C
Username

Verified customer

Submitted Oct 27 2009

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Tonsil)
Specification Tonsil
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated
Blocking step Other as blocking agent for 20 minute(s) · Concentration: 10%
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Verified customer

Submitted Mar 06 2008

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Normal Primary Fibroblast)
Specification Normal Primary Fibroblast
Fixative Paraformaldehyde
Permeabilization Yes - TritonX100
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Verified customer

Submitted Nov 05 2007

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (pathological testis)
Specification pathological testis
Fixative Formaldehyde
Antigen retrieval step Heat mediated
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1%
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Verified customer

Submitted Apr 18 2007

Thank you for your enquiry. Unfortunately we do have trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide t...

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Thank you for your your enquiry. Further to our telephone conversation I have discovered that this antibody was validated on formalin fixed sections by a 3.7% formalin fixation. Sections were permeabilised using 70% icecold methanol. 0.3% H202 was use...

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Thank you for your patience. I was away on vacation so could not reply to you earlier. From your description it appears that you are getting non-specific bands. Monoclonal Anti-Proliferating Cell Protein Ki-67 reacts with the Ki-67 nuclear antigen (3...

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We are about to implement a trial 'try risk free' scheme which says that if an antibody is purchased and used in an experiment outside that written on the datasheet, and it is found not to work, in return for the negative data (which we can add to the dat...

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