Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Knockout Validated, Immunohistochemistry, Dual RNAscope ISH-IHC Compatible, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Ki67/MKI67 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Ki67/MKI67
Asn3120-Ile3256
Accession # P46013

Specificity

Detects human Ki67/MKI67 in direct ELISAs. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse Ki67/MKI67 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Ki67/MKI67 Antibody

Ki67/MKI67 antibody in A549 Human Cell Line by Immunocytochemistry (ICC).

Ki67/MKI67 in A549 Human Cell Line.

Ki67/MKI67 was detected in immersion fixed A549 human lung carcinoma cell line using Sheep Anti-Human Ki67/MKI67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7617) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to nuclei and nucleoli. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Ki67/MKI67 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

Ki67/MKI67 in Human Breast Cancer Tissue.

Ki67/MKI67 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Sheep Anti-Human Ki67/MKI67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7617) at 1 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to the nuclei of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Ki67/MKI67 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Ki67/MKI67 at approximately 320 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Ki67/MKI67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7617) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Detection of Human Ki67/MKI67 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and Ki67 knockout HeLa cell line (KO), loaded at 0.2 mg/mL. A specific band was detected for Ki67/MKI67 at approximately 325 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human Ki67/MKI67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7617) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.
Ki67/MKI67 Antibody Specificity is Shown by Immunocytochemistry antibody in Knockout Cell Line.

Ki67/MKI67 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.

Ki67/MKI67 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line but is not detected in Ki67/MKI67 knockout (KO) HeLa cell line using Sheep Anti-Human Ki67/MKI67 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7617) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Ki-67/MKI67 in human breast cancer using Dual RNAscope®ISH and IHC.

MKi67 mRNA (red) and MKi67 protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human breast cancer. ACD’s Integrated Co-Detection Workflow was performed using ACD RNAScope Probe Hs-MKI67 (Catalog # 591771) and sheep anti-human Ki67/MKI67 polyclonal antibody (AF7617) at 5 μg/mL. Tissue was stained using RNAscope® 2.5 HD Detection Kit-RED (Catalog # 322360) and RNAscope® 2.5 LS Green Accessory Pack (Catalog # 322550). Tissue was counterstained with 50% hematoxylin (blue).

Applications for Human Ki67/MKI67 Antibody

Application
Recommended Usage

Dual RNAscope ISH-IHC Compatible

5-25 µg/mL
Sample: Formalin-fixed paraffin-embedded tissue sections of human breast cancer.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed A549 human lung carcinoma cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Knockout Validated

Ki67/MKI67 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Ki67/MKI67 knockout HeLa cell line.

Simple Western

20 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line.

Reviewed Applications

Read 2 reviews rated 5 using AF7617 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Ki67/MKI67

MKI67 (also Ki67) is a 350-400 kDa nuclear protein that belongs to a molecular group comprised of mitotic chromosome-associated proteins. Ki67 was originally recognized as an antigen associated with the monoclonal Ki67 antibody raised against Hodgkin's lymphoma nuclear material. Ki67 is contextually expressed, being potentially found in all cells that are not in the Go phase of the cell cycle. Thus, MKI67 qualifies as a cell proliferation marker. Functionally, Ki67 is known to interact with 160 kDa Hklp2, a protein that promotes centrosome separation and spindle bipolarity. It also directly interacts with NIFK, and apparently binds to UBF, thus playing a role in rRNA synthesis. Human MKI67 is 3256 amino acids (aa) in length. It contains one FHA domain (aa 8-98), followed by at least 24 utilized Ser/Thr phosphorylation sites and sixteen 120 aa repeats (aa 1000-2928) that are interspersed with at least 90 additional utilized phosphorylation sites. There are two potential isoform variants. One isoform is 315-345 kDa in size and shows a deletion of aa 136-495, while a second isoform contains a 58 aa substitution for aa 1-513. Over aa 3120-3256, human Ki67 shares 46% aa sequence identity with the mouse ortholog to Ki67.

Long Name

Antigen Identified by Monoclonal Antibody Ki67

Alternate Names

Ki-67, KIA, MIB-1, MKI67, PPP1R105, TSG126

Entrez Gene IDs

4288 (Human); 17345 (Mouse); 291234 (Rat)

Gene Symbol

MKI67

UniProt

P46013 (Human)

Additional Ki67/MKI67 Products

Product Documents for Human Ki67/MKI67 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Ki67/MKI67 Antibody

For research use only

Related Research Areas

Cancer Biomarkers
Cell Cycle

Citations for Human Ki67/MKI67 Antibody

Customer Reviews for Human Ki67/MKI67 Antibody (2)

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  • Human Ki67/MKI67 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Breast cancer tissue
    Species: Human
    Verified Customer | Posted 11/28/2024
    Human, 1:200
    Human Ki67/MKI67 Antibody AF7617
  • Name: Natalie Erdmann
    Application: Immunocytochemistry
    Sample Tested: Multiple myeloma cells line LP-1 cytospins
    Species: Human
    Verified Customer | Posted 08/24/2015
    IFF/ICC staining of LP-1 cells with AF7617
    Human Ki67/MKI67 Antibody AF7617

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