Human Ki-67/MKI67 Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human Ki-67/MKI67 in directs ELISAS.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1297A
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Human Ki-67/MKI67 synthetic peptide
    Accession # P46013
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Immunohistochemistry
    3-25 µg/mL
    See below
  • Immunocytochemistry
    0.3-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Knockout Validated
    Ki-67/MKI67 is specifically detected in Hela human cervical epithelial carcinoma parental cell line but is not detectable in Ki-67/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.
Data Examples
Immunocytochemistry
Ki-67/MKI67 in HeLa Human Cell Line.
Ki-67/MKI67 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human Ki-67/
MKI67 Monoclonal Antibody (Catalog # MAB7617) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of Ki-67/MKI67 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) either
(A) untreated or (B) treated with 5 μg/mL PHA for 5 days were stained with Rabbit Anti-Human Ki-67/MKI67 Monoclonal Antibody (Catalog # MAB7617) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # MAB1050). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Immunohistochemistry
Ki-67/MKI67 in Human Pancreatic Cancer Tissue. Ki-67/MKI67 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Rabbit Anti-Human Ki-67/MKI67 Monoclonal Antibody (Catalog # MAB7617) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Knockout Validated
Ki-67/MKI67 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.
Ki-67/MKI67 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line but is not detected in Ki-67/MKI67 knockout (KO) HeLa cell line using Rabbit Anti-Human Ki-67/MKI67 Monoclonal Antibody (Catalog # MAB7617) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • 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: Ki-67/MKI67

MKI67 (also Ki-67) is a 350-400 kDa nuclear protein that belongs to a molecular group comprised of mitotic chromosome-associated proteins. Ki-67 was originally recognized as an antigen associated with the monoclonal Ki-67 antibody raised against Hodgkin's lymphoma nuclear material. Ki-67 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, Ki-67 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 Ki-67 shares 46% aa sequence identity with the mouse ortholog to Ki-67.

  • Long Name:
    Antigen Identified by Monoclonal Antibody Ki-67
  • Entrez Gene IDs:
    4288 (Human); 17345 (Mouse); 291234 (Rat)
  • Alternate Names:
    antigen identified by monoclonal Ki-67; antigen KI-67; Ki67; Ki-67; KIA; Marker Of Proliferation Ki-67; MIB-1; MKI67; PPP1R105; proliferation-related Ki-67 antigen; Protein Phosphatase 1; Regulatory Subunit 105; TSG126
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Isotype Controls
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Normal Rabbit IgG Control

Ctrl AB-105-C 40  
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Normal Rabbit IgG Control

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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

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Rabbit IgG PE-conjugated Antibody

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Rabbit IgG APC-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

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Rabbit IgG Fluorescein-conjugated Antibody

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Goat Anti-Rabbit IgG Biotinylated Antibody

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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Rabbit IgG Antibody

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Donkey Anti-Rabbit IgG (H+L) Antibody

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