Recombinant Anti-Ki67 antibody [B56] (ab279653)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [B56] to Ki67
- Suitable for: ICC, Flow Cyt (Intra), IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Ki67 antibody [B56]
See all Ki67 primary antibodies -
Description
Mouse monoclonal [B56] to Ki67 -
Host species
Mouse -
Tested applications
Suitable for: ICC, Flow Cyt (Intra), IHC-Pmore details
Unsuitable for: WB -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human colon, ovarian carcinoma, Mouse colon, breast carcinoma, Rat colon tissues. ICC: HeLa cells. Flow Cyt: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
B56 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab279653 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC |
1/50.
|
|
| Flow Cyt (Intra) |
1/1000.
|
|
| IHC-P |
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
|
| Notes |
|---|
|
ICC
1/50. |
|
Flow Cyt (Intra)
1/1000. |
|
IHC-P
1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Required 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. -
Sequence similarities
Contains 1 FHA domain.
Contains 16 K167R repeats.
Contains 1 PP1-binding domain. -
Developmental stage
Expression 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 localization
Chromosome. 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). - Information by UniProt
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Database links
- Entrez Gene: 4288 Human
- Entrez Gene: 17345 Mouse
- Entrez Gene: 246042 Rat
- Omim: 176741 Human
- SwissProt: P46013 Human
- SwissProt: E9PVX6 Mouse
- SwissProt: Q61769 Mouse
- Unigene: 689823 Human
see all -
Alternative names
- Antigen identified by monoclonal antibody Ki 67 antibody
- Antigen identified by monoclonal antibody Ki-67 antibody
- Antigen KI-67 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab279653)
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
-
![Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [B56] (ab279653)](https://www.abcam.com/ps/products/279/ab279653/Images/ab279653-11-mouse-monoclonal-b56-to-ki67-immunocytochemistry.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Ki67 antibody [B56] (ab279653)ab279653 staining Ki67 in wild-type Hap1 cells, with negative expression in Ki67 knockout Hap1 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab279653 at 5 µg/ml and ab6046, Rabbit polyclonal to beta Tubulin at 1 µg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab279653)Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on human ovarian carcinoma. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunocytochemistry - Anti-Ki67 antibody (ab279653)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Ki67 with ab279653 at 1/50 (18.72 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar staining in HeLa cell line ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Immunocytochemistry - Anti-Ki67 antibody [B56] (ab279653)](https://www.abcam.com/ps/products/279/ab279653/Images/ab279653-459243-anti-ki67-antibody-b56-immunocytochemistry-helaKO.jpg)
Immunocytochemistry - Anti-Ki67 antibody [B56] (ab279653)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized wild-type HeLa (Human cervix adenocarcinoma epithelial cell) cells and KI67 KO HeLa cells labelling Ki67 with ab279653 at 1/50 dilution (18.72 ug/ml), followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nucleolar staining in Parental HeLa cell line and no staining in MKI67 HeLa KO cell line. ab179513 Anti-beta Tubulin rabbit monoclonal antibody at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 dilution was used to counterstain tubulin (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (ab279653)](https://www.abcam.com/ps/products/279/ab279653/Images/ab279653-7-anti-ki67-antibody-b56-flow-cytometry-hela.jpg)
Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (ab279653)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Ki67 with ab279653 at 1/1000 dilution (0.1ug)/ Right compared with a Mouse monoclonal IgG / Left isotype control. A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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![Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (ab279653)](https://www.abcam.com/ps/products/279/ab279653/Images/ab279653-8-anti-ki67-antibody-b56-flow-cytometry-parental-hela.jpg)
Flow Cytometry (Intracellular) - Anti-Ki67 antibody [B56] (ab279653)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Parental HeLa (Human cervix adenocarcinoma epithelial cells) (Right)/ MKI67 KO HeLa cells (Left) labelling Ki67 with ab279653 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab279653)Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab279653)Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on mouse breast carcinoma. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (ab279653)Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling Ki67 with ab279653 at 1/1000 dilution (0.936 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining on rat colon. The section was incubated with ab279653 for 30 mins at room temperature and followed by mouse specific IgG antibody (ab125913) for 8 mins. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Anti-Ki67 antibody (ab279653)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab279653 has been referenced in 1 publication.
- Yao J et al. lncRNA SNHG22 sponges miR-128-3p to promote the progression of colorectal cancer by upregulating E2F3. Int J Oncol 59:N/A (2021). PubMed: 34368861