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    products/primary-antibodies/ki67-antibody-sp6-ab16667.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ki67 antibody [SP6] (ab16667)

  • Datasheet
  • SDS
Reviews (125)Q&A (18)References (1744)

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Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
  • Western blot - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
  • Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)
  • Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
  • Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)
  • Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
  • Anti-Ki67 antibody [SP6] (ab16667)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP6] to Ki67
  • Suitable for: Flow Cyt (Intra), IHC-P, WB, mIHC, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free PE

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Anti-SNAP25 antibody [EPR3275] (ab109105)
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Human Ki67 ELISA Kit (ab253221)
Knockout
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Human MKI67 (Ki67) knockout HeLa cell line (ab255407)

View more associated products

Overview

  • Product name

    Anti-Ki67 antibody [SP6]
    See all Ki67 primary antibodies
  • Description

    Rabbit monoclonal [SP6] to Ki67
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), IHC-P, WB, mIHC, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Common marmoset
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Epitope

    C-terminus
  • Positive control

    • WB: HeLa cell lysate. IHC-P: Human tonsil, rat and mouse spleen tissues. ICC/IF: HeLa and HAP1 cells. Flow Cyt (intra): HAP1 cells. mIHC: Human tonsil
  • General notes

    ab16667 was switched from a hybridoma to recombinant production method on 24th October 2019.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

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. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.1% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    SP6
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Markers
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Replication
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Cancer
    • Cell cycle
    • Cell division
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Development

Associated products

  • Alternative Versions

    • Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)
    • Anti-Ki67 antibody [SP6], prediluted (ab21700)
    • Anti-Ki67 antibody [SP6] - BSA free (ab231172)
    • Alexa Fluor® 488 Anti-Ki67 antibody [SP6] (ab281847)
    • Alexa Fluor® 647 Anti-Ki67 antibody [SP6] (ab281928)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human MKI67 (Ki67) knockout HeLa cell line (ab255407)
  • KO cell lysates

    • Human MKI67 (Ki67) knockout HeLa cell lysate (ab263762)
  • Related Products

    • Ki67 peptide (ab15581)
    • Fix & Perm / Cell Fixation & Permeabilization Kit (Flow Cytometry) (ab185917)
    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • PD-L1 / PD1 Multiplex IHC-IF Antibody Panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812)
    • Anti-PCNA antibody [PC10] (ab29)
    • Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/1000.

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

IHC-P (62)
1/200.

Antigen retrieval: Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0).

Primary antibody condition: primary antibody incubation overnight at +4°C is recommended.

WB (3)
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa.
mIHC
1/200.
ICC/IF (22)
1/250.

If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min.

Notes
Flow Cyt (Intra)
1/1000.

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

IHC-P
1/200.

Antigen retrieval: Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0).

Primary antibody condition: primary antibody incubation overnight at +4°C is recommended.

WB
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa.
mIHC
1/200.
ICC/IF
1/250.

If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min.

Target

  • Function

    Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface (PubMed:27362226). Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility (PubMed:27362226). Binds DNA, with a preference for supercoiled DNA and AT-rich DNA (PubMed:10878551). Does not contribute to the internal structure of mitotic chromosomes (By similarity). May play a role in chromatin organization (PubMed:24867636). It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed.
  • Sequence similarities

    Contains 1 FHA domain.
    Contains 16 K167R repeats.
    Contains 1 PP1-binding domain.
  • Developmental stage

    Expression 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 (at protein level) (PubMed:6206131). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli (PubMed:2674163, PubMed:8799815).
  • Post-translational
    modifications

    Phosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA.
  • Cellular localization

    Chromosome. Nucleus. Nucleus, nucleolus. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the mitotic chromosome surface (PubMed:27362226). Associates with satellite DNA in G1 phase (PubMed:9510506). Binds tightly to chromatin in interphase, chromatin-binding decreases in mitosis when it associates with the surface of the condensed chromosomes (PubMed:15896774, PubMed:22002106). 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 (PubMed:22002106).
  • 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

    • Entrez Gene: 4288 Human
    • Entrez Gene: 17345 Mouse
    • Entrez Gene: 246042 Rat
    • Omim: 176741 Human
    • SwissProt: P46013 Human
    • SwissProt: E9PVX6 Mouse
    • SwissProt: Q61769 Mouse
    • Unigene: 689823 Human
    • Unigene: 80976 Human
    • Unigene: 4078 Mouse
    see all
  • 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
    • Antigen Ki67 antibody
    • KI67_HUMAN antibody
    • KIA antibody
    • Marker of proliferation Ki-67 antibody
    • MIB 1 antibody
    • MIB antibody
    • MKI67 antibody
    • PPP1R105 antibody
    • Proliferation marker protein Ki-67 antibody
    • Proliferation related Ki 67 antigen antibody
    • Protein phosphatase 1 regulatory subunit 105 antibody
    • RP11-380J17.2 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)

    ab16667 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, 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 1 hour. 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 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    This image was generated from the hybridoma version.

  • Western blot - Anti-Ki67 antibody [SP6] (ab16667)
    Western blot - Anti-Ki67 antibody [SP6] (ab16667)
    All lanes : Anti-Ki67 antibody [SP6] (ab16667) at 1/100 dilution

    Lane 1 : Ramos cell lysate
    Lane 2 : Wild-type HeLa cell lysate
    Lane 3 : MKI67 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 358 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab16667 observed at 359 kDa. Red - loading control, ab130007 observed at 125 kDa.  

     ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. ab16667 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil sections labeling Ki67 with ab16667 at 1/200 (0.156 µg/mL). 

    Image A: 
    Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer)
    Human tonsil tissue incubated with ab16667 overnight at +4°C.
    Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).
    Nuclear staining on human tonsil. The section was incubated with ab16667 overnight at +4°C.

    Image B:
    Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)
    Human tonsil tissue incubated with ab16667 on a Leica Biosystems BOND® RX instrument.
    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution2) for 20 mins.


     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)

    IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Rat spleen tissue. The section was pre-treated using heat mediated antigen retrieval with (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/mL) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on rat spleen. The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)

    IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Human tonsil tissue. The section was pre-treated using heat mediated antigen retrieval with ab93678 (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/ml) dilution and detected using ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on human tonsil is observed. The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin. 

     

  • Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)
    Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)

    Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).

    Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).

    The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.

    Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.

    DAPI (dark blue) was used as a nuclear counter stain.

    Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.

    This image was generated from the hybridoma version.

  • Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)
    Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] (ab16667)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with ab228462 at 1/100 dilution (0.52 μg/ml) (C). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    Panel A: merged staining of anti-Ki67 (magenta; Opal™690), anti-PD-L1 (red; Opal™570) and anti-PD1 (green; Opal™520) on human tonsil.
    Panel B: anti-Ki67 stained on nucleus of proliferating cells.
    Panel C: anti-PD-L1 stained on membrane of cells involved in T cell inhibition.
    Panel D: anti-PD1 stained on antigen-stimulated T cells.

    The section was incubated in three rounds of staining: in the order of ab16667 for 10 mins, ab243644 for 30 mins and ab228462 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)

    IHC image of ab16667 staining Ki67 in a section of formalin-fixed paraffin-embedded Mouse spleen tissue. The section was pre-treated using heat mediated antigen retrieval with ab93678 (citrate buffer, pH 6.0). The section was then incubated with ab16667, 1/200 (0.156 µg/mL) dilution and detected using a ready to use Goat Anti-Rabbit IgG H&L (HRP) antibody. Nuclear staining on mouse spleen.The section was incubated with ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin. 

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunofluorescent analysis of 100% methanol-fixed, None permeabilized HeLa cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in HeLa cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in parental HAP1cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

  • Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)
    Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HAP1 (Wildtype control  Human chronic myelogenous leukemia  near-haploid cell line) cells labelling Ki67 with ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730)  (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

     

  • Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)
    Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] (ab16667)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-MKI67 knockout cells (red line) stained with ab16667. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab16667,1/1000) for 30 min at 22°C. The secondary antibody used wasGoat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibodyat 1/2000 dilution for 30 min at 22°C.  A Rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MKI67 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).  Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.   This image was generated from the hybridoma version. 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)This image is courtesy of an anonymous abreview.

    Immunocytochemistry/ Immunofluorescence analysis of human cardiac stem cells labeling Ki67 with ab16667 at 1/250 dilution. Cells were fixed in paraformaldehyde and permeabilized with Triton x-100, 0.01%. Cells were blocked in BSA for 1 hour at room temperature. A polyclonal chicken anti-rabbit Alex Fluor® 488 secondary antibody was used at 1/500 dilution. 

    This image was generated from the hybridoma version.

     

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] (ab16667)Image Courtesy of Carl Hobbs, King College London, U.K.

    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% BSA 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.

    This image was generated from the hybridoma version.

     

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)
    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] (ab16667)This image is courtesy of an Abreview submitted by Peter Zentis

    ab16667 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.

    This image was generated from the hybridoma version.

     

    See Abreview

  • Anti-Ki67 antibody [SP6] (ab16667)
    Anti-Ki67 antibody [SP6] (ab16667)

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (1744)

Publishing research using ab16667? Please let us know so that we can cite the reference in this datasheet.

ab16667 has been referenced in 1744 publications.

  • Zhu X  et al. HDAC1/2 Control Proliferation and Survival in Adult Epidermis and Pre?Basal Cell Carcinoma through p16 and p53. J Invest Dermatol 142:77-87.e10 (2022). PubMed: 34284046
  • Yang W  et al. Single-Cell Transcriptomic Analysis Reveals a Hepatic Stellate Cell-Activation Roadmap and Myofibroblast Origin During Liver Fibrosis in Mice. Hepatology 74:2774-2790 (2021). PubMed: 34089528
  • de Barros Sene L  et al. Gestational Low Protein Diet Modulation on miRNA Transcriptome and Its Target During Fetal and Breastfeeding Nephrogenesis. Front Physiol 12:648056 (2021). PubMed: 34239447
  • Gao C  et al. The application of CRISPR/Cas9 system in cervical carcinogenesis. Cancer Gene Ther N/A:N/A (2021). PubMed: 34349239
  • Zhang W  et al. Downregulation of miR-335 exhibited an oncogenic effect via promoting KDM3A/YAP1 networks in clear cell renal cell carcinoma. Cancer Gene Ther N/A:N/A (2021). PubMed: 33888871
View all Publications for this product

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1-10 of 143 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph6 citrate
Permeabilization
No
Specification
Skin
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Victoria Bridgeman

Verified customer

Submitted Jun 29 2016

Immunohistochemistry free floating abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (frontal cortex)
Specification
frontal cortex
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Alexandre Magno

Verified customer

Submitted Nov 17 2022

Immunohistochemistry (Frozen sections) abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
No
Specification
brain
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: .1% · Temperature: 22°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 01 2022

Western blot abreview for Anti-Ki67 antibody [SP6]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (PANC1 cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Treatment
DHA (40 µM) 24 hrs
Specification
PANC1 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Minaam Abbas

Verified customer

Submitted Oct 25 2022

Immunohistochemistry (Frozen sections) abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (B16F10 tumors, C57BL/6J mice)
Permeabilization
Yes - 0.1% Triton X100 in PBS
Specification
B16F10 tumors, C57BL/6J mice
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 23 2022

Immunocytochemistry/ Immunofluorescence abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse lung slices)
Permeabilization
Yes - 0.5% Triton X-100 in PBS, 5min, RT
Specification
Mouse lung slices
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 14 2022

Western blot abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (CT26 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris Glycin gel)
Loading amount
40 µg
Specification
CT26 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 07 2022

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Ki67 antibody [SP6]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (B16 Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6
Permeabilization
Yes - 0.1% Triton-X100 in PBS 10'
Specification
B16 Pancreas
Blocking step
Bloxall as blocking agent for 20 minute(s) · Concentration: 100%
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 05 2022

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Ki67 antibody [SP6]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citric buffer, PH 6
Permeabilization
No
Specification
pancreas
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 27 2021

Question

Could you recommend a Ki67 antibody (clone SP6) that would work in IHC-Fr using mouse tissue?

Read More

Abcam community

Verified customer

Asked on Feb 22 2012

Answer

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

Read More

Abcam Scientific Support

Answered on Feb 22 2012

1-10 of 143 Abreviews or Q&A

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