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.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
Feature | Specification |
---|---|
Application | Cell Signaling |
Sample Volume | 16 µL |
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.
For research use only. Not for use in diagnostic procedures. All products to be used in accordance with applicable laws and regulations including without limitation, consumption and disposal requirements under European REACH regulations (EC 1907/2006).
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 Parkinsons disease (PD) and neurodegenerative disorders in general.
Application |
Cell Signaling
|
---|---|
Brand |
HTRF
|
Detection Modality |
HTRF
|
Lysis Buffer Compatibility |
Lysis Buffer 4
Lysis Buffer 5
|
Molecular Modification |
Total
|
Product Group |
Kit
|
Sample Volume |
16 µL
|
Shipping Conditions |
Shipped in Dry Ice
|
Target Class |
Phosphoproteins
|
Target Species |
Human
Mouse
|
Technology |
TR-FRET
|
Therapeutic Area |
Neuroscience
|
Unit Size |
10,000 Assay Points
|
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 proteins expression under a no-wash assay format.
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.
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.
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.
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This guide provides you an overview of HTRF applications in several therapeutic areas.
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