Revvity, 64VAV1Y74PEG, HTRF Phospho-VAV1 (Tyr174) Detection Kit, 500 Assay Points

Overview
VAV1, also known as Vav proto-oncogene 1, is a protein that plays a crucial role in cellular signaling and immune responses. It is primarily expressed in cells of hematopoietic origin, including T cells, B cells, and natural killer cells.
VAV1 acts as a guanine nucleotide exchange factor (GEF) for the Rho family GTPases, including Rac1, Rac2, and RhoA. VAV1 activates these GTPases by promoting the exchange of GDP (guanosine diphosphate) for GTP (guanosine triphosphate) on these enzymes. Through this mechanism, VAV1 is an important regulator of essential cellular processes such as cytoskeletal rearrangement, cell adhesion, proliferation, and migration.
In immune cells, VAV1 is an important intermediary in the downstream signaling of specialized antigen receptors, including T cell receptors (TCRs) and B cell receptors (BCRs). As such, VAV1 has been implicated in several diseases, including the autoimmune disorders systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). In these conditions, abnormal VAV1 signaling can contribute to immune dysregulation and inflammation. Additionally, VAV1 is involved in certain types of cancer, where aberrant activation can promote cellular transformation and metastasis.
How it works
HTRF Human Phospho-Y174 VAV1 assay principle
The Phospho-Y174 VAV1 assay measures VAV1 when phosphorylated at Tyr174. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses 2 antibodies, one labeled with a donor fluorophore and 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 independently 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.

HTRF Human Phospho-Y174 VAV1 one-plate assay protocol
Detection of Phospho-Y174 VAV1 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.

HTRF Human Phospho-Y174 VAV1 two-plate assay protocol
The two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a low volume detection plate (either HTRF 384-lv or 96-lv plate) before the addition of HTRF Phospho-Y174 VAV1 detection reagents. This protocol enables the cells' viability and confluence to be monitored. Application-Cell Signaling
Brand-HTRF
Detection Modality-HTRF
Molecular Modification-Phosphorylation
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target Class-Phosphoproteins
Technology-TR-FRET
Unit Size-500 assay points