Human Ki67 ELISA Kit (ab253221)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 21.2 pg/ml
- Range: 31.25 pg/ml - 2000 pg/ml
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Ki67 ELISA Kit
See all Ki67 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Extract 8 4.1% Inter-assay Sample n Mean SD CV% Extract 3 1.8% -
Sample type
Cell Lysate -
Assay type
Sandwich (quantitative) -
Sensitivity
21.2 pg/ml -
Range
31.25 pg/ml - 2000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell Lysate 108 101% - 116% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Ki67 ELISA Kit (ab253221) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Ki67 protein in cell lysate. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Ki67 with 21.2 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Ki67 is a nuclear marker protein that is commonly found in active proliferating cells. Encoded by the MKI67 gene, Ki67 is expressed in all phases of the cell cycle except for resting cells in G0 stage. Moreover, the presence of Ki67 protein is at its highest in the G2 phase and during mitosis. Studies have shown Ki67 to be an excellent antigen marker to determine the growth fraction of a given cell population. More particularly in cancer, which is subsequently correlated with the clinical course of action. The standard protein in ab253221 is an internal fragment comprising 7% of the full-length Ki67.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Ki67 Capture Antibody 1 x 600µl 10X Human Ki67 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPR 1 x 6ml Human Ki67 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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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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Alternative names
- Antigen identified by monoclonal antibody Ki 67
- Antigen identified by monoclonal antibody Ki-67
- Antigen KI-67
see all -
Database links
- Entrez Gene: 4288 Human
- Omim: 176741 Human
- SwissProt: P46013 Human
- Unigene: 689823 Human
- Unigene: 80976 Human
Images
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human Ki67 standard curve in 1X Cell Extraction Buffer PTR.The Ki67 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Ki67 in human A431 and Hela cell extracts based on a 250 µg/mL and 62.5 µg/mL extract load respectively.The concentrations of Ki67 were measured in duplicate and interpolated from the Ki67 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Ki67 concentration was determined to be 1,575.0 pg/mL in A431 and 1674.3 pg/mL in Hela.
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Sandwich ELISA - Human Ki67 ELISA Kit (ab253221)To learn more about the advantages of recombinant antibodies see here.
