Human Ki67 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Ki67 in Cell Lysate samples.
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Publications
- Molecules (Basel, Switzerland) 27:2022A Novel ERK2 Degrader Z734 Induces Apoptosis of MCF-7 Cells via the HERC3/p53 Signaling Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShiyao Xu,Yan Xiong,Rui Yao,Rong Tian,Zhuqing Meng,Mohamed Y Zaky,Beibei Fu,Dong Guo,Lulu Wang,Feng Lin,Xiaoyuan Lin,Haibo WuPubMed 35889210
- Biomedicines 10:2022Non-Invasive Physical Plasma Generated by a Medical Argon Plasma Device Induces the Expression of Regenerative Factors in Human Gingival Keratinocytes, Fibroblasts, and Tissue Biopsies.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBenedikt Eggers,Matthias Bernhard Stope,Jana Marciniak,Werner Götz,Alexander Mustea,James Deschner,Marjan Nokhbehsaim,Franz-Josef KramerPubMed 35453639
Key facts
- Detection method
Colorimetric
- Sample types
Cell Lysate
- Assay type
Sandwich (quantitative)
- Reactive species
Human
- Range
31.25 - 2000 pg/mL
- Assay time
1h 30m
- Sensitivity
= 21.2 pg/mL
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Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
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 (Probable).
Alternative names
Proliferation marker protein Ki-67, Antigen identified by monoclonal antibody Ki-67, Antigen KI-67, Antigen Ki67, MKI67
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Recommended products
Human Ki67 ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the quantitative measurement of Human Ki67 in Cell Lysate samples.
Alternative names
Proliferation marker protein Ki-67, Antigen identified by monoclonal antibody Ki-67, Antigen KI-67, Antigen Ki67, MKI67
Key facts
- Detection method
Colorimetric
- Sample types
Cell Lysate
- Assay type
Sandwich (quantitative)
- Reactive species
Human
- Range
31.25 - 2000 pg/mL
- Assay time
1h 30m
- Assay Platform
Pre-coated microplate (12 x 8 well strips)
- Sensitivity
= 21.2 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Extract | n 8 | mean - | SD - | C.V. 4.1 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Extract | n 3 | mean - | SD - | C.V. 1.8 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell Lysate | Average % = 108 | Range 101 - 116 % |
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
- Storage information
+4°C
Notes
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 strips
A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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.
Supplementary info
- Activity summary
Ki67 is a nuclear protein often referred to as MKI67 with a molecular weight of approximately 345 kDa. This protein is strongly associated with cell proliferation. Scientists commonly use Ki67 staining techniques to identify and quantify proliferating cells in tissues. Ki67 is expressed in the nucleus during active cell cycle phases—G1 S G2 and M—but not in resting cells in G0 phase. It is detectable through methods such as Ki67 immunofluorescence and Ki67 flow cytometry which are important for analyzing cell division.
- Biological function summary
The Ki67 protein plays an essential role in cellular proliferation processes. It does not form part of a stable complex but interacts transiently with other cell cycle-related proteins. Research indicates that Ki67 maintains the structure of the perichromosomal layer during mitosis impacting chromosome separation. It is particularly active in tissues with high cell turnover such as bone marrow and lymphoid organs.
- Pathways
Ki67 influences several key cellular pathways that control cell proliferation and differentiation. The protein acts within the regulation of the cell cycle and the PI3K/AKT signaling pathway important for cell growth and survival. Within these pathways Ki67 interacts with proteins like cyclin-dependent kinases (CDKs) which regulate transitions between different phases of the cell cycle.
- Associated diseases and disorders
Ki67 has significant implications in oncology and can be used as a biomarker for cancer prognosis. Its expression levels help determine the aggressiveness of tumors especially in breast cancer and prostate cancer. High Ki67 levels correlate with poor prognosis as it indicates rapid cell division. In cancer Ki67 associates with p53 a protein that regulates the cell cycle and function as a tumor suppressor.
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2 product images
Sandwich ELISA - Human Ki67 ELISA Kit (ab253221)
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.
Sandwich ELISA - Human Ki67 ELISA Kit (ab253221)
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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