Revvity, 64JAK2Y10PEH, HTRF Human Phospho-JAK2 (Tyr1007/1008) Detection Kit, 10,000 Assay Points

Overview
JAK2 (Janus kinase 2) belongs to the family of non-receptor Janus tyrosine kinases with JAK1, JAK3, and TYK2. A wide array of cytokines and growth factors ( IL6, IL4, IFN alpha, GM-CSF) attached to their receptors induce the phosphorylation of JAKs. The activated JAKs subsequently phosphorylate additional targets, including both the cytokine receptors and the major substrates: STATs. The JAKs/SATs signaling stimulates cell proliferation, differentiation, migration, and apoptosis. Altering JAK/Stat signaling to reduce cytokine induced pro-inflammatory responses represents an attractive target for anti-inflammatory therapies.
How it works
Phospho-JAK2 (Tyr1007/1008) assay principle
The Phospho-JAK2 (Tyr1007/1008) assay measures JAK2 when phosphorylated at Tyr1007/1008. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Phospho-JAK2 (Tyr1007/1008) 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, 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-JAK2 (Tyr1007/1008) 2-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 Phospho-JAK2(Tyr1007/1008) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 

Phospho-JAK2 (Tyr1007/1008) 1-plate assay protocol
Detection of Phosphorylated JAK2 (Tyr1007/1008) 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 4
Molecular Modification-Phosphorylation
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