Directly conjugated antibodies for immunofluorescence

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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-P, ICCmore 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.

  • EpitopeC-terminus
  • Positive control
    • Tonsil

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.
ICC/IF 1/50.
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.
ICC Use at an assay dependent concentration.

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 identified by monoclonal antibody Ki 67 antibody
    • Antigen KI-67 antibody
    • Antigen KI67 antibody
    • KI67_HUMAN antibody
    • KIA antibody
    • KIA antibody
    • MKI67 antibody
    • Proliferation related Ki 67 antigen antibody
    • Proliferation related Ki 67 antigen antibody
    • RP11-380J17.2 antibody
    see all

Anti-Ki67 antibody [SP6] images

  • 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.

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  • 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

  • Human tonsil stained with ab16667.
  • 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.

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  • 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.

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

This product has been referenced in:
  • Nishioka S  et al. The cyclin-dependent kinase inhibitor p21 is essential for the beneficial effects of renal ischemic preconditioning on renal ischemia/reperfusion injury in mice. Kidney Int 85:871-9 (2014). IHC ; Mouse . Read more (PubMed: 24336034) »
  • Dame MK  et al. Human colonic crypts in culture: segregation of immunochemical markers in normal versus adenoma-derived. Lab Invest 94:222-34 (2014). Human . Read more (PubMed: 24365748) »

See all 66 Publications for this product

Product Wall

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

Application Immunocytochemistry
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Sample Human Cell (HEK293)
Specification HEK293
Permeabilization Yes - Triton x-100, 0.01%
Fixative Formaldehyde
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Abcam user community

Verified customer

Submitted Jan 15 2015

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10 · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer, pH 6.0
Sample Mouse Tissue sections (Transgenic mouse spinal cord)
Specification Transgenic mouse spinal cord
Permeabilization Yes - Triton X-100, 0.5%
Fixative Paraformaldehyde
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Submitted Jan 08 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 0.05% trypsin
Sample Mouse Tissue sections (prostater)
Specification prostater
Permeabilization No
Fixative Formaldehyde
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Abcam user community

Verified customer

Submitted Dec 04 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
Antigen retrieval step None
Sample Human Tissue sections (human colon)
Specification human colon
Permeabilization Yes - saponin
Fixative Paraformaldehyde
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Verified customer

Submitted Dec 01 2014

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

Mr. Carl Hobbs

Verified customer

Submitted Sep 11 2014

This antibody is a tissue culture supernatent and therefore is not purified hence we have not determined its concentration.

However, typically the concentration of tissue culture supernatants is between 10-50 ug/ml so we would expect ab16667...

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Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Pituitary gland)
Specification Pituitary gland
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 31 2014

Application Immunohistochemistry (Frozen sections)
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Sample Mouse Tissue sections (Embryonic brain)
Specification Embryonic brain
Permeabilization Yes - TritonX-100
Fixative Paraformaldehyde
Username

Jianning Lu

Verified customer

Submitted Feb 20 2014

ab16667 is cultured in serum free medium.

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