The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 19520159
Use a concentration of 1.6 µg/ml.
Use at an assay dependent concentration. PubMed: 20729478
Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 359 kDa.
Use a concentration of 2 - 5 µg/ml.
Use a concentration of 0.01 - 0.1 µg/ml.
1/50 - 1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionRequired 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.
Developmental stageExpression occurs preferentially during late G1, S, G2 and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected (at protein level) (PubMed:6206131). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli (PubMed:2674163, PubMed:8799815).
Post-translational modificationsPhosphorylated. Hyperphosphorylated in mitosis (PubMed:10502411, PubMed:10653604). Hyperphosphorylated form does not bind DNA.
Cellular localizationChromosome. Nucleus. Nucleus, nucleolus. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the mitotic chromosome surface (PubMed:27362226). Associates with satellite DNA in G1 phase (PubMed:9510506). Binds tightly to chromatin in interphase, chromatin-binding decreases in mitosis when it associates with the surface of the condensed chromosomes (PubMed:15896774, PubMed:22002106). Predominantly localized in the G1 phase in the perinucleolar region, in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix (PubMed:22002106).
Antigen identified by monoclonal antibody Ki 67 antibody
Antigen identified by monoclonal antibody Ki-67 antibody
Antigen KI-67 antibody
Antigen KI67 antibody
Antigen Ki67 antibody
Marker of proliferation Ki-67 antibody
MIB 1 antibody
Proliferation marker protein Ki-67 antibody
Proliferation related Ki 67 antigen antibody
Protein phosphatase 1 regulatory subunit 105 antibody
Anti-Ki67 antibody images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab66155)This image is courtesy of an abreview submitted by Virginia Gutiérrez
Immunohistochemical analysis of formaldehyde fixed mouse skin sections labelling Ki67 with ab66155 at a dilution of 1:1000. The secondary antibody used was a goat anti-rabbit IgG (HRP). Antigen retrieval was performed using Citrate buffer pH 6.0 for 20 minutes at 97ºC.
Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody (ab66155)This image is courtesy of an Abreview submitted by Francesco Elia Marino
ab66155 staining Ki67 in human LnCAP cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with ethanol and triton, and blocked for 1 hour at 26°C. Samples were incubated with primary antibody (1/200 in blocking buffer) for 16 hours at 4°C. An undiluted IRDye® 800CW-conjugated goat anti-rabbit IgG (H+L) polyclonal was used as the secondary antibody.
Immunohistochemistry (Frozen sections) - Anti-Ki67 antibody (ab66155)Image courtesy of an anonymous Abreview.
ab66155 staining Ki67 in murine brain tissue by Immunohistochemistry (Frozen sections). Tissue was fixed with paraformaldehyde and then blocked with 5% serum for 1 hour at 25°C followed by incubation with the primary antibody at a 1/200 dilution for 12 hours at 4°C. ab96919 Donkey polyclonal to anti-rabbit DyLight® 488 (IgG - H&L), pre-adsorbed was used as the secondary antibody undiluted.
Immunohistochemistry (Frozen sections) - Anti-Ki67 antibody (ab66155)This image is courtesy of an Abreview submitted by Elizandra Braganhol
ab66155 at 1/300 dilution staining Ki67 in rat glioma tissue by immunohistochemistry (frozen sections). Sections were paraformaldehyde fixed prior to blocking in 1% BSA for 30 minutes at 20°C and then incubated with ab66155 for 18 hours at 4°C.
This image shows a human lymph node stained with ab66155 at 1/200 dilution for 10 min at RT.
Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody (ab66155)This image is courtesy of an anonymous Abreview.
ab66155 staining ki67 in proliferating mouse astrocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldahyde, permeabilized with 0.3% TritonX 100 and 3M Glycine and blocked with 10% BSA for 20 minutes at 20°C. Samples were incubated with primary antibody (1/250 in 4% BSA/PBS) for 18 hours at 4°C. ab150073 at a dilution of 1/500 was used as the secondary antibody.
Astrocytes (orange) labelled with Ki67 proliferation marker (violet) and counter stained with DAPI (blue).
Scale bar 20um, magnification x40 oil.
References for Anti-Ki67 antibody (ab66155)
This product has been referenced in:
Su B et al. Diallyl disulfide suppresses epithelial-mesenchymal transition, invasion and proliferation by downregulation of LIMK1 in gastric cancer. Oncotarget7:10498-512 (2016).
Read more (PubMed: 26871290) »
Huang M et al. Roles of limbal microvascular net and limbal stroma in regulating maintenance of limbal epithelial stem cells. Cell Tissue Res359:547-63 (2015).
Read more (PubMed: 25398719) »