Rabbit Monoclonal Anti-Ki67 Antibody from Abcam

Abcam’s rabbit monoclonal Anti-Ki67 antibody is an excellent protein loading control antibody.

  • Product Name - Anti-Ki67 antibody [SP6]
  • Description - Rabbit monoclonal [SP6] to Ki67
  • Host Species - Rabbit

Specificity

Many Abcam’s customers have been successful in using ab16667 on rat and mouse for IHC. However, Abcam is presently enhancing their testing for these species and thus cannot guarantee that this product will reliably work on rat and mouse in IHC.

Abcam does not guarantee western blot for rat and mouse.

  • Tested Applications - Suitable for IHC-FoFr, ICC/IF, Flow Cyt, IHC-Fr, WB, and IHC-P
  • Species Reactivity - Reacts with rat, mouse, common marmoset, human
  • Immunogen - Synthetic peptide within Human Ki67 aa 1200-1300. The precise sequence is proprietary.
  • Epitope - C-terminus

Positive Control

  • IHC-P: Human tonsil and testis tissue
  • Rat esophagus, liver tissue, and small intestine
  • Common marmoset spleen tissue
  • Mouse embryonic skin tissue
  • Rat lymph node tissue
  • Transgenic mouse spinal cord tissue
  • ICC/IF: HAP1 cells
  • HEp-2 cells
  • Flow Cyt: HAP1 cells
  • Human cardiac stem cells
  • Rat cardiomyocytes

Applications

Abcam’s Abpromise gurantee covers the application of ab16667 in the tested applications listed below. Note: Recommended starting dilutions are included in the application notes; ideal dilutions/concentrations should be established by the end user.

Application Abreviews Notes
IHC-FoFr ☆☆☆☆☆ 1/5000.
Antigen retrieval: Boil tissue section in 10 mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at room temperature for 20 minutes.
ICC/IF ☆☆☆☆☆ 1/250.
Flow Cyt ☆☆☆☆☆ 1/1000.
ab172730, Rabbit monoclonal isotype, is ideal for use as an isotype control along with this antibody.
IHC-Fr ☆☆☆☆☆ 1/1000. A higher dilution is recommended for frozen tissues than for FFPE tissues. Abcam suggests a starting dilution of 1/500.
WB Can be used at an assay dependent concentration. PubMed: 20562294
Mouse and rat species are suggested based on IHC results, Abcam does not assure western blot for rat and mouse.
IHC-P ☆☆☆☆☆ 1/200

Target

Function

The rabbit monoclonal Anti-Ki67 antibody is needed to maintain each mitotic chromosome spread in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). It associates with the mitotic chromosome surface—the perichromosomal layer—and encompasses a considerable fraction of the chromosome surface (PubMed:27362226).

The antibody stops chromosomes from disintegrating into a single chromatin mass by developing a steric and electrostatic charge barrier: the protein has a high net electrical charge and serves as a surfactant, dispersing chromosomes and facilitating independent chromosome motility (PubMed:27362226). It binds DNA, with an affinity for supercoiled DNA and AT-rich DNA (PubMed:10878551). It does not add to the internal structure of mitotic chromosomes (by similarity).

The rabbit monoclonal Anti-Ki67 antibody may have a role to play in chromatin organization (PubMed:24867636). However, it is uncertain whether it has a direct impact in chromatin organization or whether it is an indirect result of its function in keeping mitotic chromosomes dispersed.

Sequence Similarities

  • Comprises 1 FHA domain
  • Comprises 1 PP1-binding domain
  • Comprises 16 K167R repeats

Developmental Stage

Expression takes place preferentially during late G1, S, G2, and M phases of the cell cycle, while in cells in G0 phase, the antigen cannot be noticed (at protein level) (PubMed:6206131). The antigen is present at maximum level in G2 phase and during mitosis (at protein level). In interphase, it develops fiber-like structures in fibrillarin-deficient regions adjoining nucleoli (PubMed:2674163, PubMed:8799815).

Post-Translational Modifications

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

Cellular Localization

Chromosome. Nucleus. Nucleus, nucleolus. Binds to the surface of the mitotic chromosome—the perichromosomal layer—and encompasses a considerable fraction of the mitotic chromosome surface (PubMed:27362226). Binds to satellite DNA in G1 phase (PubMed:9510506). Binds securely to chromatin in interphase, chromatin-binding reduces in mitosis when it adheres to the surface of the condensed chromosomes (PubMed:22002106, PubMed:15896774).

Mainly localized in the G1 phase in the perinucleolar region, and in the later phases, it is also noticed all over the nuclear interior, being mainly localized in the nuclear matrix (PubMed:22002106).