Recombinant Anti-Ki67 antibody [SP6], prediluted (ab21700)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-31031-anti-ki67-antibody-sp6-prediluted-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [SP6] to Ki67, prediluted
- Suitable for: WB, ICC, Flow Cyt, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Overview
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Product name
Anti-Ki67 antibody [SP6], prediluted
See all Ki67 primary antibodies -
Description
Rabbit monoclonal [SP6] to Ki67, prediluted -
Host species
Rabbit -
Specificity
ab21700 recognises Ki67.
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Tested applications
Suitable for: WB, ICC, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Ramos and HeLa cell lysates. IHC-P: Human tonsil and thymus tissue; Human endometrial and breast adenocarcinoma tissue. ICC: HeLa and HAP1 cells. Flow Cyt:: HAP1 cells.
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General notes
FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
This product was switched from hybridoma to recombinant format on 25th October 2019.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: Tris buffered saline, 1% BSA
Inert stabilizer -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
SP6 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab21700 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 |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa.
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| ICC |
Use at an assay dependent concentration.
Use at an assay dependent concentration. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min |
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| Flow Cyt |
Use at an assay dependent concentration.
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| IHC-P | (2) |
1/1.
Antigen retrieval: heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) |
| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 358 kDa. |
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ICC
Use at an assay dependent concentration. Use at an assay dependent concentration. If fixing cells in 4% PFA (20 min, room temp), it is recommended to permeabilized cells with 0.1% Triton-X for 5 min |
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Flow Cyt
Use at an assay dependent concentration. |
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IHC-P
1/1. Antigen retrieval: heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) |
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 [SP6], prediluted (ab21700)Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human tonsil tissue, staining Ki67 with ab21700.
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![Western blot - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-386662-anti-ki67-antibody-sp6-prediluted-western-blot-human-lysate.jpg)
Western blot - Anti-Ki67 antibody [SP6], prediluted (ab21700)All lanes : Anti-Ki67 antibody [SP6] (ab16667) at 1/100 dilution
Lane 1 : Ramos cell lysate
Lane 2 : Wild-type HeLa cell lysate
Lane 3 : MKI67 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 358 kDa
Observed band size: 359 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab16667).
Lanes 1 - 3: Merged signal (red and green). Green - ab16667 observed at 359 kDa. Red - loading control, ab130007 observed at 125 kDa.
ab16667 was shown to react with Ki67 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255407 (knockout cell lysate ab263762) was used. Wild-type and Ki67 knockout samples were subjected to SDS-PAGE. ab16667 and Anti-Vinculin antibody [VIN-54] (ab130007) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-7-anti-ki67-antibody-sp6-immunocytochemistry-parental-hap1.jpg)
Immunocytochemistry - Anti-Ki67 antibody [SP6], prediluted (ab21700)This data was developed using ab16667, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 100% methanol-fixed, None permeabilized parental HAP1 cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in parental HAP1cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
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![Immunocytochemistry - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-6-anti-ki67-antibody-sp6-immunocytochemistry-hela.jpg)
Immunocytochemistry - Anti-Ki67 antibody [SP6], prediluted (ab21700)This data was developed using ab16667, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 100% methanol-fixed, None permeabilized HeLa cells labelling Ki67 with ab16667 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing nucleolar staining in HeLa cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 dilution (2 µg/mL).
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![Flow Cytometry - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-8-anti-ki67-antibody-sp6-flow-cytometry-parental-hap1.jpg)
Flow Cytometry - Anti-Ki67 antibody [SP6], prediluted (ab21700)This data was developed using ab16667, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HAP1 (Wildtype control Human chronic myelogenous leukemia near-haploid cell line) cells labelling Ki67 with ab16667 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-31030-anti-ki67-antibody-sp6-prediluted-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human endometrial adenocarcinoma tissue, staining Ki67 with ab21700. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-31029-anti-ki67-antibody-sp6-prediluted-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human thymus tissue, staining Ki67 with ab21700. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-31028-anti-ki67-antibody-sp6-prediluted-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody [SP6], prediluted (ab21700)Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human breast adenocarcinoma tissue, staining Ki67 with ab21700. -
![Anti-Ki67 antibody [SP6], prediluted (ab21700)](https://www.abcam.com/ps/products/21/ab21700/Images/ab21700-9-benefits-of-recombinant-antibodies.png)
Anti-Ki67 antibody [SP6], prediluted (ab21700)
Datasheets and documents
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SDS download
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Datasheet download
References (18)
ab21700 has been referenced in 18 publications.
- Zhang S et al. Hsa_circ_0007494 suppresses prostate cancer progression via miR-616/PTEN axis. Exp Cell Res 395:112233 (2020). PubMed: 32810511
- Lou Q et al. miR-448 targets IDO1 and regulates CD8+ T cell response in human colon cancer. J Immunother Cancer 7:210 (2019). PubMed: 31391111
- Pan F et al. Dietary riboflavin deficiency promotes N-nitrosomethylbenzylamine-induced esophageal tumorigenesis in rats by inducing chronic inflammation. Am J Cancer Res 9:2469-2481 (2019). PubMed: 31815047
- Kefayat A et al. C-phycocyanin: a natural product with radiosensitizing property for enhancement of colon cancer radiation therapy efficacy through inhibition of COX-2 expression. Sci Rep 9:19161 (2019). PubMed: 31844085
- Al Asmari AK & Khan AQ Investigation of in vivo potential of scorpion venom against skin tumorigenesis in mice via targeting markers associated with cancer development. Drug Des Devel Ther 10:3387-3397 (2016). PubMed: 27799739