Overview

  • Product nameAnti-Ki67 antibody [SP6]
    See all Ki67 primary antibodies
  • Description
    Rabbit monoclonal [SP6] to Ki67
  • Tested applicationsIHC-FoFr, ICC/IF, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Marmoset (common)
  • Immunogen

    Synthetic peptide within Human Ki67 aa 2300-2400 (C terminal). The exact sequence is proprietary.
    Database link: P46013

  • EpitopeC-terminus
  • Positive control
    • Tonsil ICC/IF: HAP1 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab16667 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 1/5000.

Antigen retrieval: Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min.

ICC/IF 1/250.
IHC-Fr 1/1000. A higher dilution is recommended for frozen tissues than for FFPE tissues. We suggest a starting dilution of 1/500.
WB Use at an assay dependent concentration. PubMed: 20562294
IHC-P 1/100.

Target

Anti-Ki67 antibody [SP6] images

  • ab16667 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 ab16667 at 1/250 dilution 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.

  • ab16667 staining Ki67 in Rat liver tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 2 hours at 22°C. Samples were incubated with primary antibody (1/500 in PBS-T + 1% BSA) for 18 hours at 4°C. A Biotin-conjugated goat anti-rabbit IgG monoclonal (1/2000) was used as the secondary antibody.

    See Abreview

  • Immunohistochemical analysis of Human Tonsil tissue labeling Ki-67 with ab16667 at 1:200. The HRP/AEC-staining procedure was used for detection.

  • ab16667 staining Ki67 in human testis by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% B.S.A. for 10 minutes at 21ºC followed by incubation with the primary antibody for 2 hours at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

    See Abreview

  • ab16667 staining Ki67 in mouse embryonic skin tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% B.S.A. for 10 minutes at 21ºC followed by incubation with the primary antibody for 16 hours at 1/50. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

    See Abreview

  • ab16667 staining Ki67 in Marmoset (common) spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% B.S.A. for 10 minutes at 21ºC followed by incubation with the primary antibody for 2 hours at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

    See Abreview

  • ab16667 staining Ki67 in rat oesophagus by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Samples were then blocked with 1% B.S.A. for 10 minutes at 21ºC followed by incubation with the primary antibody for 30 minutes at 1/100. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/250 dilution.

    See Abreview

  • ab16667 staining Ki67 in rat small intestine tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using 10mM citrate buffer pH 6.0. Samples were then blocked with 10% serum for 20 minutes at room temperature followed by incubation with the undiluted primary antibody for 30 minutes. A biotin-conjugated goat anti-rabbit polyclonal was used as secondary antibody at a 1/2000 dilution.

    See Abreview

  • ab166667 staining Ki67 - Proliferation Marker in Human HEp-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with Triton X-100 0.25% in PBS. Samples were incubated with primary antibody (1/50 in DPBS) for 1 hour at 21°C. An Atto488-conjugated Donkey anti-rabbit polyclonal (1/50) was used as the secondary antibody.

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  • ab16667 staining Ki67 (red) in transgenic mouse spinal cord tissue sections (depelted of oligodendrocytes) by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.5% Triton X-100 and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in 10mM citrate buffer, pH 6, for 20 minutes at 97°C in a water bath. The sample was incubated with primary antibody (1/300 in PBS + 0.1% Triton X-100 + 1% serum) at 25°C for 16 hours. An Alexa Fluor® 594-conjugated donkey anti-rabbit IgG (H+L) polyclonal (1/700) was used as the secondary antibody. Counterstained with Iba1 (green) a marker for microglia and DAPI.

    See Abreview

References for Anti-Ki67 antibody [SP6] (ab16667)

This product has been referenced in:
  • Chen Y  et al. FGFR antagonist induces protective autophagy in FGFR1-amplified breast cancer cell. Biochem Biophys Res Commun N/A:N/A (2016). Read more (PubMed: 26993162) »
  • Siopi E  et al. Anxiety- and Depression-Like States Lead to Pronounced Olfactory Deficits and Impaired Adult Neurogenesis in Mice. J Neurosci 36:518-31 (2016). Read more (PubMed: 26758842) »

See all 97 Publications for this product

Product Wall

Either ab16667 or ab27619 would work for your chosen application and species.

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Ear)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization No
Specification Ear
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 12 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: pH6, Citrate based
Permeabilization No
Specification colon
Blocking step Serum Free Protein Block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
Fixative 10% NBF
Username

Dr. Faye Doherty

Verified customer

Submitted Apr 21 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Cat Tissue sections (liver)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: tris-EDTA, pH9.0
Permeabilization No
Specification liver
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 13 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Cartilage)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Permeabilization Yes - Wash Buffer from Dako with Tween
Specification Cartilage
Fixative Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

Submitted Mar 14 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Spleen)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Permeabilization Yes - Wash Buffer from Dako with Tween
Specification Spleen
Fixative Paraformaldehyde
Username

Mr. Rudolf Jung

Verified customer

Submitted Mar 14 2016

Application Immunohistochemistry (Frozen sections)
Sample Dog Tissue sections (gut)
Permeabilization No
Specification gut
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Acetone
Username

Abcam user community

Verified customer

Submitted Jan 28 2016

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Hippocampus)
Permeabilization Yes - TritonX-100
Specification Hippocampus
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Acetone
Username

Miss.

Verified customer

Submitted Jan 20 2016

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (bain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification bain
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 19 2016

Application Flow Cytometry
Sample Human Cell (human esophageal cancer cell lines)
Permeabilization Yes - 90% methanol, -20oC
Gating Strategy viable cell population was gated based on a stably expressed fluorescent marker (iRFP670)
Specification human esophageal cancer cell lines
Username

Ms. Tatiana Karakasheva

Verified customer

Submitted Nov 09 2015

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