Total LRRK2 cellular kit
All-in-one kit for robust detection of Total LRRK2
Phospho-LRRK2 (Ser935) cellular kit HTRF®
This HTRF kit enables the cell-based quantitative detection of LRRK2 protein phosphorylated at Serine 935 directly in your cell type of interest.
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Low sample consumption
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No-wash
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Validated on PBMC
Overview
The phospho-LRRK2 (Ser935) kit measures endogenous LRRK2 protein phosphorylated at Ser935 directly in your cell type of interest. LRRK2 is a significant target in neuroscience research involving Parkinsons disease (PD) and neurodegenerative disorders in general.
Benefits
- VALIDATED ON PATIENT PBMC'S
- SPECIFICITY
Phospho-LRRK2 (Ser935) assay principle
The phospho-LRRK2 (Ser935) assay measures LRRK2 when phosphorylated at Ser935. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The phospho-LRRK2 (Ser935) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is 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 proteins phosphorylation state under a no-wash assay format.
Phospho-LRRK2 (Ser935) 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 phospho-LRRK2 (Ser935) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Phospho-LRRK2 (Ser935) 1-plate assay protocol
Detection of phosphorylated LRRK2 (Ser935) 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 phospho-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 phospho-assay is 8-fold more sensitive than the Western Blot: only 2,500 cells are sufficient for minimal signal detection when using the HTRF phospho- LRRK2 assay while 20,000 cells are needed for a Western Blot signal.
Phospho LRRK2 detection in NIH 3T3 cells
Cells were cultured in 6-well plates and treated with increasing concentrations of CZC
and GSK
for 2 hours. Medium was removed and replaced with Cisbio lysis buffer for 1 hour. 16 µL of lysate were taken from each well and transferred to a 384-well plate for detection with HTRF reagents. The Cisbio phosphorylated LRRK2 assay enabled clear determination of both inhibitors IC50s, in agreement with literature.
Simplified pathway dissection with HTRF phospho-assays and CyBi-felix liquid handling
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
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
Cell Signaling: Biomarkers, Phospho- & total-protein assays - Flyers
PI3K/AKT/mTor translational control pathway - Posters
Universal HTRF® phospho-protein platform: from 2D, 3D, primary cells to patient derived tumor cells
Analysis of a large panel of diverse biological samples and cellular models - Posters
From 2D, 3D cell cultures to xenografts: A smart HTRF platform to maximize anticancer drug discovery
One technology across all samples - Application Notes
HTRF phospho assays reveal subtle drug induced effects in tumor-xenografts
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
HTRF cell-based phospho-protein data normalization
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
HTRF phospho-total lysis buffer: a universal alternative to RIPA lysis buffers
Increased flexibility of phospho-assays - Application Notes
Exploring LRRK2 phosphorylation with HTRF® assays
LRRK2 kit, a game changer in neuroscience - Posters
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
Best practices for analyzing brain samples with HTRF® phospho assays for neurosciences
Insider Tips for successful sample treatment - Technical Notes
HTRF Alpha-tubulin Housekeeping kit
Properly interpret your compound effect - Application Notes
Optimize your HTRF cell signaling assays on tissues
HTRF and WB compatible guidelines - Technical Notes
Key guidelines to successful cell signaling experiments
Mastering the art of cell signaling assays optimization - Guides
HTRF phospho-assays reveal subtle drug-induced effects
Detailed protocol and direct comparison with WB - Posters
A single technology for 2D cells, 3D cells, and xenograft models - Posters
Best practices for analyzing tumor xenografts with HTRF phospho assays
Protocol for tumor xenograft analysis with HTRF - Technical Notes
Detection of cellular total and phospho LRRK2 with HTRF® assays in BIOMIMESYS® 3D culture
In collaboration with Celenys - Posters
How to run a cell based phospho HTRF assay
What to expect at the bench - Videos
Unleash the potential of your phosphorylation research with HTRF
Unmatched ease of use, sensitivity and specificity assays - Videos
Cell-based kinase assays in HTS ? potential and limitations for primary and secondary screening
In collaboration with Bayer - Scientific Presentations
Product Insert LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Product Insert
Product Insert LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Product Insert
HTRF Product Catalog 2020 July update
All your HTRF assays in one document! - Catalog
A guide to Homogeneous Time Resolved Fluorescence
General principles of HTRF - Guides
How HTRF compares to Western Blot and ELISA
Get the brochure about technology comparison. - Brochures
HTRF® cell signaling platform combined with iCell® Hepatocytes
A solution for phospho-protein analysis in metabolic disorders - Posters
Unleash the potential of your phosphorylation research with HTRF
A fun video introducing you to phosphorylation assays with HTRF - Videos
How to run a cell based phospho HTRF assay
3' video to set up your Phospho assay - Videos
Molecular basis of neuroinflammation and neurodegeneration diseases
The essential guide for extending your knowledge on the molecular mechanisms of neurodegenerative diseases - Guides
Neurodegeneration and its main related diseases
Discover this infographic design on neurodegenerative diseases - Infographics
Safety Data Sheet LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (DEU) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (ELL) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (FRA-FR) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (ITA) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (SPA) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (ENG-GB) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet (ENG-US) LLRK2-S935 Kit / 6FLRKPEG
6FLRKPEG - Safety Data Sheet
Safety Data Sheet LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (DEU) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (ELL) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (FRA-FR) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (ITA) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (SPA) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (ENG-GB) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (ENG-US) LLRK2-S935 Kit / 6FLRKPEH
6FLRKPEH - Safety Data Sheet
Safety Data Sheet (DEU) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (ELL) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (FRA-FR) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (ITA) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (SPA) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (ENG-GB) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
Safety Data Sheet (ENG-US) LLRK2-S935 Kit / 6FLRKPEY
6FLRKPEY - Safety Data Sheet
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