Revvity, 64CREPEH, HTRF Human and Mouse Phospho-CREB (Ser133) Detection Kit, 10,000 Assay Points

Overview
The phospho-CREB (S133) kit is optimal for the quantitative detection of the cyclic AMP-responsive element-binding protein when phosphorylated at S133, thus enabling the phosphorylation status of CREB to be checked with respect to its steady-state level in cells. CREB functions as a transcription factor, signaling downstream from MAPK/ERK and activated by several pathways, such as MAPK/ERK, PI3K/AKT and stress signaling. CREB plays a critical role in promoting neuronal plasticity, cognitive abilities, cell proliferation, survival, and glucose homeostasis.
How it works
Phospho-CREB (S133) assay principle
The Phospho-CREB (S133) assay measures CREB when phosphorylated at S133. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-CREB (S133) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, 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 proteins phosphorylation state under a no-wash assay format.

 

Phospho-CREB (S133) 2-plate assay protocol
The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-CREB (S133) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Phospho-CREB (S133) 1-plate assay protocol
Detection of Phosphorylated CREB (S133) 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
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-Metabolism/Diabetes NASH/Fibrosis Oncology & Inflammation
Unit Size-10,000 assay points