Revvity, 64JAK3TPEH, HTRF Human Total JAK3 Detection Kit, 10,000 Assay Points

Overview
The Human Total JAK3 detection assay monitors cellular JAK3 protein levels.
The Janus family kinases JAK1, JAK2, JAK3, and TYK2 are non-receptor protein tyrosine kinases that signal downstream cytokine receptors (mainly IL-R and IFN-R) and activate the STAT transcription factors. A specific JAK/JAK combination associates with a specific receptor and gives specificity to the downstream JAK/STAT signaling pathway. JAK3, in combination with JAK1, associates exclusively with the IL-2Rγ family receptors, and triggers the activation of the transcription factors STAT1, STAT3, STAT5, and STAT6 to regulate lymphocyte maturation, survival, activation, and differentiation.
JAK3 dysregulation has been reported in cancer and immune disorders. The main therapeutic strategies consist in developing potent and selective inhibitors/degraders of JAK3.
How it works
Total JAK3 assay principle
The Total JAK3 assay quantifies the expression level of JAK3 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total JAK3 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 the presence of JAK3 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 JAK3 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 Total JAK3 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Total JAK3 one-plate assay protocol
Detection of Total JAK3 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 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
Therapeutic Area-Inflammation
Unit Size-10,000 assay points