Revvity, 64HP21PEH, HTRF Human Total p21 Detection Kit, 10,000 Assay Points

Overview
p21, also known as cyclin-dependent kinase inhibitor 1 or CDK-interacting protein 1, is a key cyclin-CDK complex inhibitor that binds to cyclin-CK2, cyclin-CDK1, and cyclin-CDK4/6, and regulates their activity. p21 works in tandem with p53 to achieve downstream cell-cycle arrest following DNA damage, and is negatively regulated by ubiquitin-ligase-driven protein degradation.
HTRF assays offer many advantages over other technologies:

Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
HTRF Human p21 assay principle
The HTRF Human p21 assay quantifies the expression level of human p21 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses two labeled antibodies: one coupled to a donor fluorophore, the other to an acceptor. In presence of p21 in a cell lysate, the addition of these conjugates brings the donor fluorophore into close proximity with the acceptor, and thereby generates a FRET signal. Its intensity is directly proportional to the concentration of the protein present in the sample, and provides a means of assessing the protein’s expression under a no-wash assay format.

 

HTRF Human p21 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of human p21 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored. Application-Cell Signaling
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 2
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Therapeutic Area-Oncology & Inflammation
Unit Size-10,000 assay points