Product DescriptionMouse anti-Ki-67 Monoclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Application DetailsWestern blotting (1:1,000-1:5,000) and Immunocytochemistry (1:2,000-1:5,000). Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
TargetKi-67
SpecificityHuman,reacts with Human only. Does not react with Mouse or Rat.
Target Host SpeciesHuman
Species ReactivityHuman
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1
Clone Name6B4
ConjugateUnconjugated
Immunogen DescriptionRecombinant human Ki-67 protein (amino acids 1,111-1,490) expressed in and purified from E. coli.
Purity DescriptionProtein G purified
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, and sodium azide
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute with 100 µL sterile-filtered, ultrapure water to achieve a 1 mg/mL concentration. Centrifuge to remove any insoluble material.
Storage InstructionsStore lyophilized antibody at 2-8°C. After reconstitution divide into aliquots and store at -20°C for long-term storage. Store at 2-8°C short-term (up to 4 weeks) with an appropriate antibacterial agent. Avoid repetitive freeze/thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundRequired to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly (PubMed:27362226). Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface (PubMed:27362226). Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility (PubMed:27362226). Binds DNA, with a preference for supercoiled DNA and AT-rich DNA (PubMed:10878551). Does not contribute to the internal structure of mitotic chromosomes (By similarity). May play a role in chromatin organization (PubMed:24867636). It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed (Probable). Ref: uniprot.org
Left: Immunofluorescence analysis of HeLa cells costained with mouse antibody to Ki-67 (1:2,000, red) and chicken antibody to vimentin (C-1409-50, 1:10,000, green). Ki-67 protein is predominantly localized in nucleoli of _cells in interphase, while in mitotic cells Ki-67 is associated with condensed chromosome. Ki-67 is not expressed in cells in quiescent G0 state. Blue: DAPI staining of nuclear DNA. Right: Western blot analysis of equal amounts of HeLa (Lane 2) and HEK293 (Lane 3) cell lysates probed with mouse antibody to Ki-67 (1:2,000, Lanes 2 and 3). Two bands above 250 kDa are observed, corresponding to major Ki-67 isoforms of molecular weight ~345 and 395 kDa.
Analysis of Ki67 expression (green) in cultured HeLa cells with mouse anti-Ki67 antibody by Immunocytochemistry. The nuclei of rapidly dividing cells have clumps of Ki67 associated with nucleoli, while nearby quiescent cells express little or no Ki67. Cells are counterstained with chicken anti-lamin A/C antibody (C-1698-100, red) which reveals the nuclear lamina. Blue: DAPI nuclear stain.
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