Revvity, 64RSK1S0PEH, HTRF Human and Mouse Phospho-RSK1 (Ser380) Detection Kit, 10,000 Assay Points

Overview
The Phospho-RSK1 (S380) Cellular Kit is designed for the robust quantification of phosphorylated RSK1 directly using a streamlined mix-and-read, no-wash protocol. RSK1 plays a pivotal role in cell proliferation and differentiation. This kit can be used from basic research to high throughput drug screening. Its versatility means it is suitable for many applications, ranging from basic research to the analysis of pharmacological questions in cellular models.
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
Phospho-RSK1 (S380) assay principle
The Phospho-RSK1 (S380) Assay measures RSK1 phosphorylation at Serine 380. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total RSK1 assay uses 2 labeled antibodies: one with a donor fluorophore, the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein’s phosphorylation state under a no-wash assay format.

 

Phospho-RSK1 (S380) 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis. Then lysates are transferred into a 384-well low volume detection plate before the addition of HTRF Phospho-RSK1 (S380) detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Phospho-RSK1 (S380) 1-plate assay protocol
Detection of Phospho-RSK1 (S380) 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
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 2
Molecular Modification-Phosphorylation
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-Infectious Diseases Metabolism/Diabetes Oncology & Inflammation Rare Diseases
Unit Size-10,000 assay points