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Total LRRK2 cellular kit HTRF®

This HTRF kit detects cellular LRKK2 modulation in the study of Parkinson's Disease It can be used as a normalization assay with the phospho-LRRK2 kit.

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  • Low sample consumption Low sample consumption
  • No-wash No-wash
  • Validated on PBMC Validated on PBMC

This HTRF kit detects cellular LRKK2 modulation in the study of Parkinson's Disease It can be used as a normalization assay with the phospho-LRRK2 kit.

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Overview

This assay is designed to monitor the modulation of LRKK2 protein levels and for use as a normalization assay with the phospho-LRRK2 kits. LRRK2 is a significant target in neuroscience research involving Parkinson’s disease (PD) and neurodegenerative disorders in general.

Benefits

  • VALIDATED ON PATIENT PBMC'S
  • SPECIFICITY

Total LRRK2 assay principle

The total LRRK2 assay quantifies the expression level of LRRK2 in a cell lysate. Contrary to 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, the other to an acceptor. Both antibodies are highly specific for a distinct epitope on the protein. In 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 assay principle

Total LRRK2 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 total LRRK2 HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Total-LRRK2 2-plate assay protocol

Total LRRK2 1-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 enables miniaturization while maintaining robust HTRF quality.
Total-LRRK2 1-plate assay protocol

HTRF total LRRK2 assay compared to Western Blot

Murine NIH-3T3 cells were cultured for 2 days until reaching 100% confluency. Following cell lysis, soluble fractions were then collected after a 10 min centrifugation. Serial dilutions of the cell lysate were performed in the supplemented lysis buffer and 16 µL of each dilution were dispensed and analyzed side-by-side by WB and by HTRF. The HTRF total assay is 2-fold more sensitive than the Western Blot: only 10,000 cells are sufficient for minimal signal detection when using the HTRF total LRRK2 assay while 20,000 cells are needed for a Western Blot.
HTRF total LRRK2 assay compared to Western Blot

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Open R&D: Sanofi Access Platform

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Cisbio lysis buffer compatibility

Cell Signaling: Biomarkers, Phospho- & total-protein Assays - Flyers

HTRF cellular phospho-protein assays

Physiologically relevant results fo fast flowing research - Flyers

Species compatibility

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Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells

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Valuable guidelines for efficiently analyzing and interpreting results - Application Notes

HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers

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Key guidelines to successful cell signaling experiments

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HTRF Product Catalog 2020 July update

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

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How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

HTRF® cell signaling platform combined with iCell® Hepatocytes

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Neurodegeneration and its main related diseases

Discover this infographic design on neurodegenerative diseases - Infographics

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

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