Revvity, 64GSPT1TPEH, HTRF Human Total GSPT1 Detection Kit, 10,000 Assay Points

Overview
The GSPT1 (G1 to S phase transition 1) protein, primarily known for its role in protein synthesis termination, has emerged as a hot target in drug discovery This is especially the case in Targeted Protein Degradation, as elevated levels of GSPT1 have been observed in certain cancers, including breast, lung, and ovarian cancers, as well as Acute Myeloid Lymphoma. Furthermore, GSPT1 may contribute to chemotherapy resistance in cancer cells. Its overexpression can lead to decreased sensitivity to anticancer drugs, making it a potential therapeutic target to overcome resistance.
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 GSPT1 assay principle
The HTRF Total GSPT1 assay quantifies the expression level of GSPT1 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total GSPT1 assay uses two labeled antibodies, one coupled to a donor fluorophore, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In presence of GSPT1 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 GSPT1 two-plate assay protocol
The 2-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 the Total GSPT1 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Human Total GSPT1 one-plate assay protocol
Detection of Total GSPT1 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
Lysis Buffer Compatibility-Lysis Buffer 1 Lysis Buffer 3 Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Target Species-Human
Technology-TR-FRET
Unit Size-10,000 assay points