Anti-Ki67 antibody - Proliferation Marker (ab15580)
- Product nameAnti-Ki67 antibody - Proliferation MarkerSee all Ki67 primary antibodies ...
- DescriptionRabbit polyclonal to Ki67 - Proliferation Marker
- Tested applicationsIHC - Wholemount, IHC-P, IHC-FrFl, Flow Cyt, IHC-Fr, ICC/IF, ICC, WB, IHC-FoFr more details
- Species reactivityReacts with: Mouse, Rat, Horse, Cow, Dog, Human, Indian Muntjac, Monkey, Chinese Hamster, Syrian Hamster
Synthetic peptide conjugated to KLH derived from within residues 1200 - 1300 of Human Ki67.
(Peptide available as ab15581.)
- Positive controlWB: Hela whole cell lysate IF: mouse (P0) olfactory bulb, MEF1 IHC-Fr: mouse P7 brain sections IHC-P: mouse spleen
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Polyclonal
- Research Areas
Our Abpromise guarantee covers the use of ab15580 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC - Wholemount||IHC - Wmt: Use at an assay dependent concentration.|
|IHC-P||IHC-P: Use a concentration of 0.1 - 10 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|IHC-FrFl||IHC-FrFl: 1/500. (see Abreview)|
|Flow Cyt||Flow Cyt: 1/100.|
|IHC-Fr||IHC-Fr: 1/100 - 1/1000.|
|ICC/IF||ICC/IF: 1/100 - 1/1000.|
|ICC||ICC: Use at an assay dependent dilution.|
|WB||WB: Use a concentration of 1 µg/ml. Detects a band of approximately 345, 395 kDa (predicted molecular weight: 359 kDa).Can be blocked with Ki67 peptide (ab15581).|
|IHC-FoFr||IHC-FoFr: Use at an assay dependent dilution.|
- FunctionThought to be required for maintaining cell proliferation.
- Sequence similaritiesContains 1 FHA domain.
- Developmental stageExpression of this antigen 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.
- Cellular localizationNucleus. Nucleus > nucleolus. Chromosome. 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. In mitosis, it is present on all chromosomes.
- Entrez Gene: 513220 Cow
- Entrez Gene: 100686578 Dog
- Entrez Gene: 100055048 Horse
- Entrez Gene: 4288 Human
- Entrez Gene: 17345 Mouse
- Entrez Gene: 246042 Rat
- Omim: 176741 Human
- SwissProt: Q3SZM1 Cow
- SwissProt: P46013 Human
- SwissProt: Q91VE6 Mouse
- SwissProt: Q5RJM0 Rat
- Unigene: 689823 Human
- Unigene: 80976 Human
- Unigene: 4078 Mouse
- Unigene: 233802 Rat
- Antigen identified by monoclonal antibody Ki 67 antibodyAntigen identified by monoclonal antibody Ki 67 antibodyAntigen KI-67 antibody
- Antigen KI67 antibodyKI67_HUMAN antibodyKIA antibodyKIA antibodyMKI67 antibodyProliferation related Ki 67 antigen antibodyProliferation related Ki 67 antigen antibodyRP11-380J17.2 antibody
Anti-Ki67 antibody - Proliferation Marker images
All lanes : Anti-Ki67 antibody - Proliferation Marker (ab15580) at 1 µg/ml
Lane 1 : Hela whole cell lysate
Lane 2 : Hela whole cell lysate with
Ki67 peptide (ab15581) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size : 359 kDa
Fluorescent confocal microscopy (20x) of mouse (P0) olfactory bulb, outer glomeruli layer, showing Ki67 immunoreactivity (ab15580; 1/1000; overnight at RT, 0.25% TX-100 no blocking step) using a secondary goat anti-rabbit fluorescent antibody (Alexa Fluor 488;1/300 2h at RT.
ab15580 staining Ki67 in SK-N-SH cells treated with NADA (N-Arachidonyldopamine) (ab120099), by ICC/IF. Decrease in Ki67 expression correlates with increased concentration of NADA (N-Arachidonyldopamine), as described in literature.
The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120099 (NADA (N-Arachidonyldopamine)) in ethanol, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab15580 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
SK-N-SH cells were permitted to grow to confluency, then serum starved for 48 hours and predominantly driven into G0. The cells were then paraformaldehyde fixed and immunofluorescently labelled with anti-Ki67 (ab15580) at a dilution of 1/1000. The majority of the cells show little or no Ki67 staining, indicating they are in G0 arrest (red cells). Two cells however show strong nucleolar Ki67 staining indicating they are still cycling (green cells). The DNA is stained with DAPI and is shown in red. The Ki67 staining is shown in green. x 63 magnification.
Similar results were seen with an asynchronous population of HeLa cells. The Ki67 staining was localised to the periphery of the nucleoli and throughout the nucleoplasm of proliferating cells. (This data is not shown but is available upon request).
Immunostaining for Rabbit polyclonal to Ki67 - Proliferation Marker (ab15580) on Mouse P7 brain (frozen) sections. Sections were fixed with paraformaldehyde, neither a blocking nor antigen retrieval steps were included in this protocol. Secondary antibody used was Rabbit IgG antibody (ab7082; 1/250).
ab15580 at 1/50 staining Human umbilical artery endothelial cells by ICC/IF. The tissue was paraformaldehyde fixed and blocked before permeabilization with saponin and incubation with the antibody for 16 hours. A FITC conjugated goat anti-rabbit IgG was used as the secondary. The merged image shows those cells expressing Ki67 from the total number of exponential cells.
IHC image of ab15580 stained human skin carcinoma FFPE section. Section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30 seconds at 125°C. Section was incubated with ab15580 at a dilution of 1:200 for 1h at room temperature and detected using an HRP conjugated polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ab15580 staining Ki67-Proliferation Maker in human colon tissue sections by IHC-P (formaldehyde-fixed paraffin-embedded sections). Tissue samples were fixed with formaldehyde and blocked with 10% goat serum for 30 minutes at 25°C. Antigen retrieval was by heat mediation in Target Retrieval Solution. Samples were incubated with primary antibody 1/5000 (TBST) for 1 hour at 25°C.
ICC/IF image of ab15580 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15580, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ICC/IF image of ab15580 stained MEF1 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15580, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab15580 staining in mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab15580, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab15580 staining Ki67 - Proliferation Marker in Human PBMCs by Flow Cytometry. Cells were isolated by Ficoll density separation, fixed in paraformaldehyde and permeabilized in 0.1% Saponin + PBS. The sample was incubated with the primary antibody (1/100 in PBS + 1% FCS and 0.09% sodium azide) for 1 hour at 4°C. A phycoerythrin-conjugated Goat anti-rabbit IgG (1/100) was used as the secondary antibody.
Gating Strategy: Lymphocytes and Monocytes
1 = isotype; 2 = PBMCs untreated; 3 = PBMCs treated with PHA.
ab15580 staining Ki67 - Proliferation Marker in Human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 4% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in a citrate buffer (pH 6.0). Samples were incubated with primary antibody (5 µg/ml in blocking buffer) for 16 hours at 4°C. A Texas Red ® Goat anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.
References for Anti-Ki67 antibody - Proliferation Marker (ab15580)
This product has been referenced in:
- Egea L et al. GM-CSF produced by nonhematopoietic cells is required for early epithelial cell proliferation and repair of injured colonic mucosa. J Immunol 190:1702-13 (2013). Mouse . Read more (PubMed: 23325885) »
- Feldman DE et al. Pluripotency factor-mediated expression of the leptin receptor (OB-R) links obesity to oncogenesis through tumor-initiating stem cells. Proc Natl Acad Sci U S A 109:829-34 (2012). Read more (PubMed: 22207628) »