Anti-Mouse Ki67 Monoclonal Antibody () for Flow Cytometry (Intracellular) - E9PVX6

Core Product Specifications and Parameters

Biological Background: Ki67 Function and Localization

• Ki67, encoded by the Mki67 gene, is a large proliferation marker protein with a molecular weight of ~351 kDa (350864 Da from UniProt) that associates with mitotic chromosomes. It acts as a chromosome repellent during early mitosis and a chromosome attractant during late mitosis, ensuring proper chromosome dispersion and clustering (PubMed:27362226). Related references: PMID:27362226
• During early mitosis, Ki67 relocalizes from nucleoli to the chromosome surface, forming extended brush-like structures. These structures provide a steric and electrostatic barrier, acting as a surfactant to prevent chromosome collapse and enable independent motility (PubMed:27362226). Related references: PMID:27362226
• In anaphase, dephosphorylation of Ki67 and exposure of a conserved basic patch trigger RNA-dependent liquid-liquid phase separation on the chromosome surface, promoting chromosome clustering and facilitating nuclear reassembly. This phase transition switches Ki67 from a repellent to an attractant.
• Ki67 binds DNA with a preference for supercoiled and AT-rich DNA, and also binds premature ribosomal RNAs during anaphase, which contributes to phase separation. However, it does not contribute to the internal structure of mitotic chromosomes (PubMed:26949251). Related references: PMID:26949251
• Subcellularly, Ki67 localizes to the nucleolus during interphase and translocates to the chromosome surface during mitosis. It is predominantly expressed in proliferating cells, serving as a reliable marker for cell cycle activation.
• Post-translational modifications include acetylation, phosphorylation, and ubiquitination, which regulate its mitotic functions and dynamic localization.

Experimental Guidance and Technical Tips

• For intracellular flow cytometry, use a permeabilization method compatible with nuclear antigens, such as methanol or saponin-based buffers, to ensure antibody access to Ki67 in the nucleolus and on chromosomes.
• Titrate the antibody between 0.1–1 µg per 10^6 cells to determine the optimal staining concentration for your specific cell type and protocol.
• Include a positive control, such as proliferating mouse cell lines (e.g., NIH/3T3, CTLL-2) or stimulated primary lymphocytes, and a negative control (e.g., isotype control or resting cells) to validate staining specificity.
• Fix cells promptly to preserve Ki67 distribution; prolonged storage may affect epitope integrity. Paraformaldehyde fixation followed by permeabilization is commonly used.
• The conjugate is compatible with standard red laser (633–647 nm) detection; ensure appropriate filter sets and minimal compensation if multiplexing.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio is your dedicated partner for IVD raw-material sourcing, offering a comprehensive portfolio from concept to clinic. This anti-Ki67 antibody is part of our range of cell proliferation and oncology markers, supported by our ability to supply validated antibody pairs, optimized recombinant proteins, and ancillary reagents. Whether you need bulk monoclonal antibodies or customized raw-material solutions for diagnostic development, contact CamelBio for reliable, one-stop support.