Revvity, 64AP2TPEB, HTRF Human and Mouse Total AP2 Detection Kit, 1,000 Assay Points

Overview
The Total AP2 cellular assay monitors total AP2 beta, and is used to detect the expression of endogenous or overexpressed AP2 beta in various cells. AP2 beta are components of the adaptor protein complex 2 (AP-2) and are known to n as cargo proteins. They play several roles in cells, including the internalization of GPCRs through a clathrin-dependent endocytosis process by direct interaction with beta arrestins. ons of AP2 beta ion cause specific dampening of cellular responses to stimuli such as hormones, neurotransmitters, or sensory signals. As AP2 essed in various tissues, it is believed that AP2 may play a major role in many cancers or other human diseases (diabetes) by ing receptor-mediated ns.
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
Total AP2 assay principle
The Total AP2 assay quantifies the expression level of AP2 beta in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total AP2 assay uses two labeled antibodies, one coupled to a donor fluorophore and the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of AP2 in a cell extract, 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.

 

Total AP2 two-plate assay protocol
Detection of total AP2 ​​​​​​​ with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.

 

Total AP2 one-plate assay protocol
Detection of total AP2 with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Cardiovascular Inflammation Metabolism/Diabetes NASH/Fibrosis
Unit Size-1,000 assay points