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Knockout Tested Rabbit Recombinant Monoclonal Ki67 antibody. Carrier free. Suitable for ICC/IF, WB, mIHC, IHC-P, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 2 publications.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (AB197547), expandable thumbnail
  • Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (AB197547), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (AB197547), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (AB197547), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (AB197547), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFWBmIHCIHC-PFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Expected
Tested
Expected
Rat
Expected
Expected
Expected
Tested
Expected
Common marmoset
Predicted
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

-

Notes

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

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/100

Notes

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.

Species

Rat

Dilution info

1/100

Notes

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.

Species

Human

Dilution info

1/100

Notes

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.

Predicted
Predicted

Species

Common marmoset

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Common marmoset

Dilution info

-

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

4 products for Alternative Version

Target data

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 (Probable).

Alternative names

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal Ki67 antibody. Carrier free. Suitable for ICC/IF, WB, mIHC, IHC-P, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 2 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

SP6

Purity

Tissue culture supernatant

Specificity

Ki67 is mainly expressed in proliferating cells. For normal tissue samples (e.g., liver, kidney), no staining may be typically observed due to low level of proliferation and little expression of Ki67. For malignant tissue samples (e.g., colon carcinoma, breast carcinoma), it is more easily to find Ki67 in the proliferating cells of these tissues (PMID: 6206131, 10653597, 34183782).

FURTHER INFORMATION ON SPECIFICITY <a href="https://www.abcam.com/ps/products/197/ab197547/documents/Anti-Ki67-antibody-SP6-BSA-and-Azide-free-FURTHER-INFORMATION-ON-SPECIFICITY-Chinese-v1a-ab197547 (website).pdf">(Chinese Version)</a>

Epitope

C-terminus

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab197547 is the carrier-free version of Anti-Ki67 antibody [SP6] ab16667.

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 batch production

For more information, read more on recombinant antibodies.

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

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Ki67 is a nuclear protein often referred to as MKI67 with a molecular weight of approximately 345 kDa. This protein is strongly associated with cell proliferation. Scientists commonly use Ki67 staining techniques to identify and quantify proliferating cells in tissues. Ki67 is expressed in the nucleus during active cell cycle phases—G1 S G2 and M—but not in resting cells in G0 phase. It is detectable through methods such as Ki67 immunofluorescence and Ki67 flow cytometry which are important for analyzing cell division.

Biological function summary

The Ki67 protein plays an essential role in cellular proliferation processes. It does not form part of a stable complex but interacts transiently with other cell cycle-related proteins. Research indicates that Ki67 maintains the structure of the perichromosomal layer during mitosis impacting chromosome separation. It is particularly active in tissues with high cell turnover such as bone marrow and lymphoid organs.

Pathways

Ki67 influences several key cellular pathways that control cell proliferation and differentiation. The protein acts within the regulation of the cell cycle and the PI3K/AKT signaling pathway important for cell growth and survival. Within these pathways Ki67 interacts with proteins like cyclin-dependent kinases (CDKs) which regulate transitions between different phases of the cell cycle.

Associated diseases and disorders

Ki67 has significant implications in oncology and can be used as a biomarker for cancer prognosis. Its expression levels help determine the aggressiveness of tumors especially in breast cancer and prostate cancer. High Ki67 levels correlate with poor prognosis as it indicates rapid cell division. In cancer Ki67 associates with p53 a protein that regulates the cell cycle and function as a tumor suppressor.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

15 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Ki67 immunohistochemistry staining of human tonsil using rabbit anti-Ki67 antibody

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).
    Immunohistochemical analysis of formalin-fixed paraffin-embedded human tonsil sections labeling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/200 dilution (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 Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C.
    Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer pH 6.0).
    Nuclear staining on human tonsil. The section was incubated with Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C.

    Image B:
    Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection)
    Human tonsil tissue incubated with Anti-Ki67 antibody [SP6] ab16667 on a Leica Biosystems BOND® RX instrument.
    Heat mediated antigen retrieval with Citrate buffer (pH 6.0 epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    Lanes 1-3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.

    Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa. Loss of signal was observed when knockout sample Human MKI67 (Ki67) knockout HeLa cell lysate ab263762 was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody VIN-54] (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) at 1/100 dilution

    Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6] - BSA free (Anti-Ki67 antibody [SP6] - BSA free ab231172) at 1/100 dilution

    Lanes 1 - 3: Western blot - Anti-Ki67 antibody [SP6], prediluted (Anti-Ki67 antibody [SP6], prediluted ab21700) at 1/100 dilution

    Lane 1: Ramos cell lysate at 20 µg

    Lane 2: Wild-type HeLa cell lysate at 20 µg

    Lane 3: MKI67 knockout HeLa cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 358 kDa

    Observed band size: 124 kDa, 359 kDa

    Lanes 1 - 3: Merged signal (red and green). Green - Anti-Ki67 antibody [SP6] ab16667 observed at 359 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007 observed at 125 kDa.

    Anti-Ki67 antibody [SP6] ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human MKI67 (Ki67) knockout HeLa cell line ab255407 (knockout cell lysate Human MKI67 (Ki67) knockout HeLa cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. Anti-Ki67 antibody [SP6] ab16667 and Anti-Vinculin antibody [VIN-54] (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    IHC image of Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Ki67 immunofluorescence staining of HeLa cells using rabbit anti-Ki67 antibody

    This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.
    Immunofluorescent analysis of 100% methanol-fixed None permeabilized HeLa cells labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    IHC image of Anti-Ki67 antibody [SP6] 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 Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The section was then incubated with Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).

  • Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Ki67 flow cytometry staining of HAP1 cells using rabbit anti-Ki67 antibody

    Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HAP1 (Wildtype controlHuman chronic myelogenous leukemianear-haploid cell line) cells labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    IHC image of Anti-Ki67 antibody [SP6] 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 Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The section was then incubated with Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] ab16667 overnight at +4°C. DAB was used as the chromogen. The section was then counterstained with haematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    Anti-Ki67 antibody [SP6] 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 Anti-Ki67 antibody [SP6] ab16667 at 1/250 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 (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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    Anti-Ki67 antibody [SP6] 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% BSA for 10 minutes at 21°C 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.

    This image was generated from the hybridoma version.

  • Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Multiplex immunohistochemistry - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using Anti-Ki67 antibody [SP6] ab16667, the same antibody clone in a different buffer formulation.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling PD1 with Anti-PD1 antibody [EPR23119-111] ab243644 at 1/500 dilution (1.02 µg/mL) (D), Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/200 dilution (0.15 μg/ml) (B) and PD-L1 with Anti-PD-L1 antibody [SP142] - C-terminal 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 Anti-Ki67 antibody [SP6] ab16667 for 10 mins, Anti-PD1 antibody [EPR23119-111] ab243644 for 30 mins and Anti-PD-L1 antibody [SP142] - C-terminal 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.

  • Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Western blot - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    This data was developed using the same antibody clone in a different buffer formulation (abAB16667).

    Western blot: Anti-MKI67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) staining at 1/500 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Ki67 antibody [SP6] ab16667 was shown to bind specifically to MKI67. A band was observed at 359 kDa in wild-type A549 cell lysates with no signal observed at this size in MKI67 knockout cell line. To generate this image, wild-type and MKI67 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-Ki67 antibody [SP6] (Anti-Ki67 antibody [SP6] ab16667) at 1/500 dilution

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: MKI67 knockout A549 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

    Observed band size: 359 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil Ki67 with ab197547 at a concentration of 5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab197547 anti Ki67 antibody was incubated at 37oC for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Tissue Microarrays stained for "Anti-Ki67 antibody [SP6]” using "Anti-Ki67 antibody [SP6] ab16667" at 1/200 dilution (0.145 μg/ml) in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Citrate Buffer) ab93678 (citrate buffer, pH 6.0). The sections were incubated with Anti-Ki67 antibody [SP6] ab16667 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer) (Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880). Hematoxylin was used as the counter stain.

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6] - BSA and Azide free (ab197547)

    Ki67 immunohistochemistry staining of human tonsil using rabbit anti-Ki67 antibody

    This data was developed using the same antibody clone in a different buffer formulation (Anti-Ki67 antibody [SP6] ab16667).

    Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling Ki67 with Anti-Ki67 antibody [SP6] ab16667 at 1/500 dilution. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). Anti-Ki67 antibody [SP6] ab16667 anti Ki67 antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

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