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HTRF Human Total MET Detection Kit, 500 Assay Points
HTRF Human Total MET Detection Kit, 500 Assay Points
Shipping box for Revvity reagent kits
The HTRF Total MET detection kit is designed to monitor the expression level of cellular MET.
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 |
The HTRF Total MET detection kit is designed to monitor the expression level of cellular MET.
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).
Product Variants
Unit Size: 500 Assay Points
Part #:
64METTPEG
List Price
USD 2,147.00
Unit Size: 10,000 Assay Points
Part #:
64METTPEH
List Price
USD 12,490.00
HTRF Human Total MET Detection Kit, 500 Assay Points
Shipping box for Revvity reagent kits
HTRF Human Total MET Detection Kit, 500 Assay Points
Product information
Overview
c-MET, also called tyrosine-protein kinase MET or hepatocyte growth factor receptor (HGFR), is a human MET gene-coded protein with tyrosine kinase activity. It acts as a tyrosine kinase receptor, with functions related to wound healing, organogenesis, and embryonic development.
MET is expressed in epithelial cells and is triggered by hepatocyte factors HGF and HSF, with ramifications into various pathways like RAS-MAPK, AKT, and STAT3 signaling.
Abnormal activation of MET is linked to cancer progression, with effects on tumor growth, angiogenesis, and tumor cell migration & invasion. These relationships between MET and tumor cells are found across multiple cancer types (liver, brain, breast, kidney, etc) and are suspected to originate from a hijacking of MET and its ligand expression regulation, which leads to their overexpression and autocrine activation of MET.
Specifications
| Application |
Cell Signaling
|
|---|---|
| Brand |
HTRF
|
| Detection Modality |
HTRF
|
| Lysis Buffer Compatibility |
Lysis Buffer 1
Lysis Buffer 2
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Sample Volume |
16 µL
|
| Shipping Conditions |
Shipped in Dry Ice
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
TR-FRET
|
| Unit Size |
500 Assay Points
|
Video gallery
HTRF Human Total MET Detection Kit, 500 Assay Points
HTRF Human Total MET Detection Kit, 500 Assay Points
How it works
Total MET assay principle
The HTRF Total MET assay quantifies the expression level of MET in a cell lysate. Unlike Western Blot, the assay is entirely plate-based, and does not require gels, electrophoresis, or transfer. The Total MET 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 MET 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 MET 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 Total MET detection reagents. This protocol enables the cells' viability and confluence to be monitored.
Total MET one-plate assay protocol
Detection of Total MET 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.
Assay validation
Validation of Phospho & Total MET detection in HGF-stimulated A431 cells
A431 cells were plated under 10µL in a 96-well plate (100,000 cells/well) in complete culture medium, and incubated overnight at 37ºC, 5% CO2. The next day, medium was removed and 50µL of complete medium was added before a 2h incubation at 37ºC, 5% CO2. After the incubation, the cells were treated with 50µL of increasing concentrations of human HGF diluted in complete medium for 15 minutes at 37ºC, 5% CO2. After medium removal, cells were then lysed with 60 µL of lysis buffer #2 and phospho-total protein blocking reagent for 45 minutes at RT under gentle shaking. After lysis, 16µL of lysate were transferred into a low volume white microplate before the addition of 2µL each of the donor and acceptor HTRF Phospho-MET (Y1234/1235) or Total MET detection reagents. The HTRF signal was recorded after an overnight incubation.
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