Revvity, 64KI67PEH, HTRF Human Ki-67 Detection Kit, 10,000 Assay Points

Overview
The expression of the proliferation related nuclear antigen Ki-67, also known as Ki67 or MKI67, is a cellular marker for proliferation which is only detected in dividing cells. Ki-67 is a non-histone protein characterized as a nuclear matrix-associated proliferation-related antigen present throughout the cell cycle, but not in resting, quiescent, cells (G0 phase). Ki-67 is widely used in routine pathology as a proliferation marker to measure the growth fraction of cells in human tumors. The Ki-67 protein present in the nucleus of proliferating cells is released upon cell lysis, and the Ki-67 protein extracted in the lysate is quantified using our Ki-67 HTRF assay. The Ki-67 quantitation assay is an alternative to BrdU or EdU incorporation assays, which are exogenous proliferation markers incorporated into the newly synthesized DNA of dividing cells. The scope of Ki-67 is similar to that of the proliferating cell nuclear antigen (PCNA).
How it works
Assay principle
​The Ki-67 quantification kit is based on a sandwich format involving two specific antibodies, respectively labelled with Cryptate (donor) and d2 (acceptor). The Ki-67 protein contained in the proliferating cell nucleus and extracted in cell lysates binds to monoclonal anti-Ki67 Eu Cryptate and monoclonal anti-Ki67 d2 conjugates. When the dyes are in close proximity, the excitation of the donor with a light source (laser or flash lamp) triggers a Fluorescence Resonance Energy Transfer (FRET) towards the acceptor, which in turn fluoresces at a specific wavelength (665 nm). The specific signal modulates positively in proportion to Ki-67 concentration.

 

Assay protocol
​The assay protocol, using a 384-well small volume white plate or a Cisbio low volume 96-well plate (20 µL final), is described on the right. 4 µL of sample (lysate) + 12 µL of lysis buffer, or 16 µL of standard are dispensed directly into the plate for detection by HTRF reagents. The antibodies labelled with HTRF donor and acceptor can be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay can be run in 96- to 384-well plates by simply resizing each addition volume proportionally. Application-Protein Quantification
Brand-HTRF
Detection Modality-HTRF
Product Group-Kit
Sample Volume-4 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Biomarkers
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points