Abcam, ab253221, Human Ki67 ELISA Kit

Size: 1 x 96Tests
Human Ki67 ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Ki67 with a sensitivity of 21.2 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step. - Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader - Design your own immunoassay: we also offer the conjugation-ready antibody pair
Key facts
Detection method:Colorimetric,
Sample types:Cell Lysate,
Reacts with:Human,
Assay type:Sandwich (quantitative),
Sensitivity:= 21.2 pg/mL,
Range:31.25 - 2000 pg/mL,
Assay time:1h 30m,
Assay Platform:Pre-coated microplate (12 x 8 well strips)

Product details:
Human Ki67 ELISA Kit ab253221 is a rapid single-wash 90-min Sandwich ELISA to measure Human Ki67 in cell lysate. This SimpleStep sensitivity is 21.2 pg/mL.
How the assay works
Human Ki67 SimpleStep ELISA
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.
Assay Specificity
Our SimpleStep ELISA
kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.
Human Ki67 ELISA Kit ab253221 protocol summary
1. Mix: add samples/standards to the wells together with the capture and detector antibody cocktail. Incubate 1 hr at room temperature
2. Wash
3. Add TMB development solution - incubate for 10 min
4. Add Stop solution
5. Read the results on a plate reader at 450 nm
Design your own immunoassay
We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:
- Anti-Ki67 antibody [EPR20582-139] - BSA and Azide free (Capture)
ab252656
- Anti-Ki67 antibody [EPR20582-7] - BSA and Azide free (Detector)
ab252656
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.
REACH authorisation
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.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-+4°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
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.
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.