Human Ki67/MKI67 Antibody

Catalog # Availability Size / Price Qty
AF7617
AF7617-SP
Ki67/MKI67 in A549 Human Cell Line.
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Product Details
Citations (5)
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Human Ki67/MKI67 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Ki67/MKI67 in direct ELISAs. In direct ELISAs, approximately 10% cross-reactivity with recombinant mouse Ki67/MKI67 is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Ki67/MKI67
Asn3120-Ile3256
Accession # P46013
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Simple Western
20 µg/mL
HeLa human cervical epithelial carcinoma cell line and MCF‑7 human breast cancer cell line.
Immunohistochemistry
5-15 µg/mL
See below
Dual RNAscope ISH-IHC
5-25 µg/mL
Formalin-fixed paraffin-embedded tissue sections of human breast cancer.
Immunocytochemistry
5-15 µg/mL
See below
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.
 

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

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

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.

Immunohistochemistry Ki67/MKI67 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western View Larger

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.

Knockout Validated View Larger

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.

Knockout Validated Ki67/MKI67 Antibody Specificity is Shown by Immunocytochemistry antibody in Knockout Cell Line. View Larger

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.

Dual RNAscope ISH-IHC Compatible View Larger

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).

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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
Entrez Gene IDs
4288 (Human); 17345 (Mouse); 291234 (Rat)
Alternate Names
antigen identified by monoclonal Ki-67; Antigen Ki67; antigen KI-67; Ki67; Ki-67; KIA; Marker Of Proliferation Ki-67; MIB-; MIB-1; MKI67; PPP1R105; Proliferation Marker Protein Ki-67; proliferation-related Ki-67 antigen; Protein Phosphatase 1; Regulatory Subunit 105; TSG126

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Citations for Human Ki67/MKI67 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Multiple cancer types rapidly escape from multiple MAPK inhibitors to generate mutagenesis-prone subpopulations
    Authors: TE Hoffman, C Yang, V Nangia, CR Ill, SL Spencer
    bioRxiv : the preprint server for biology, 2023-03-21;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Tau promotes oxidative stress-associated cycling neurons in S phase as a pro-survival mechanism: possible implication for Alzheimer's disease
    Authors: M Denechaud, S Geurs, T Comptdaer, S Bégard, A Garcia-Núñ, LA Pechereau, T Bouillet, Y Vermeiren, PP De Deyn, R Perbet, V Deramecour, CA Maurage, M Vanderhaeg, S Vanuytven, B Lefebvre, E Bogaert, N Déglon, T Voet, M Colin, L Buée, B Dermaut, MC Galas
    Progress in neurobiology, 2022-12-06;0(0):102386.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  3. Fibroblastic FAP promotes intrahepatic cholangiocarcinoma growth via MDSCs recruitment
    Authors: Y Lin, B Li, X Yang, Q Cai, W Liu, M Tian, H Luo, W Yin, Y Song, Y Shi, R He
    Neoplasia, 2019-11-20;21(12):1133-1142.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  4. B-cell lymphoma 2 is associated with advanced tumor grade and clinical stage, and reduced overall survival in young Chinese patients with colorectal carcinoma
    Authors: J Wang, G He, Q Yang, L Bai, B Jian, Q Li, Z Li
    Oncol Lett, 2018-04-13;15(6):9009-9016.
    Species: Human
    Sample Types: Tissue Homogenates, Whole Tissue
    Applications: IHC-P, Western Blot
  5. Amino Acid Transporter Slc38a5 Controls Glucagon Receptor Inhibition-Induced Pancreatic ? Cell Hyperplasia in Mice
    Authors: J Kim, H Okamoto, Z Huang, G Anguiano, S Chen, Q Liu, K Cavino, Y Xin, E Na, R Hamid, J Lee, B Zambrowicz, R Unger, AJ Murphy, Y Xu, GD Yancopoulo, WH Li, J Gromada
    Cell Metab., 2017-06-06;25(6):1348-1361.e8.
    Species: Human
    Sample Types:

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