Revvity, 64LRRKTPEH, HTRF Human and Mouse Total LRRK2 Detection Kit, 10,000 assay points

Overview
Leucine-Rich Repeat Kinase 2 (LRRK2) is a multi-domain protein that phosphorylates a subset of Rab GTPases. The LRRK2/Rab pathway operates at the interface of vesicular trafficking, lysosomal functioning, and autophagy, and thus plays a crucial role in immune response and neuroinflammation. Ser1292 autophosphorylation serves as an indicator of LRRK2 kinase activity. Pathogenic LRRK2 mutations, such as the common mutation G2019S, increase LRRK2 kinase activity, resulting in a toxic hyperactive protein that contributes to the Parkinson’s Disease (PD) phenotype. Consequently, LRRK2 inhibitors and degraders are considered promising therapeutic agents.
How it works
Total LRRK2 assay principle

The Total LRRK2 assay quantifies the expression level of LRRK2 in a cell lysate. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The Total LRRK2 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 LRRK2 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 LRRK2 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 HTRF Total LRRK2 detection reagents. This protocol enables the cells' viability and confluence to be monitored.

 
Total LRRK2 one-plate assay protocol

Detection of Total LRRK2 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 allows miniaturization while maintaining robust HTRF quality. Application-Cell Signaling
Automation Compatible-Yes
Brand-HTRF
Detection Modality-HTRF
Lysis Buffer Compatibility-Lysis Buffer 4
Molecular Modification-Total
Product Group-Kit
Sample Volume-16 µL
Shipping Conditions-Shipped in Dry Ice
Target-LRRK2
Target Class-Phosphoproteins
Target Species-Human Mouse
Technology-TR-FRET
Therapeutic Area-Neuroscience
Unit Size-10,000 assay points