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| Feature | Specification |
|---|---|
| Application | Cell Signaling |
| Sample Volume | 10 µL |
Epidermal growth factor receptor (EGFR, EGF Receptor, Erb-1, or HER1) is a transmembrane protein and receptor for members of the epidermal growth factor (EGF) family. Binding of EGFR ligands drives receptor homodimerization or hetero-dimerization, leading to the activation of the EGFR tyrosine kinase domain and specific tyrosine residues. The signals transmitted from the EGF receptor to the nucleus lead to the regulation of various biological functions, such as cell proliferation, differentiation, survival, adhesion, migration, and angiogenesis. Mutations in EGFR, including mutation L858R in exon 21, result in EGFR overexpression and are associated with cancer.
The AlphaLISA SureFire Human Total EGF Receptor L858R Detection Kit is a sandwich immunoassay for the quantitative detection of total EGF Receptor L858R in cellular lysates, using Alpha Technology.
The Total-AlphaLISA SureFire Ultra assay measures the expression level of a protein target in a cell lysate.
The Total-AlphaLISA SureFire Ultra assay uses two antibodies which recognize two different distal epitopes on the targeted protein. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the assay specific antibody, labeled with a CaptSure tag. Donor beads are coated with streptavidin to capture one of the detection antibodies, which is biotinylated. In the presence of targeted protein, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, allowing the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of protein present in the sample.
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well OptiPlate™ plate before the addition of Total-AlphaLISA SureFire Ultra detection reagents. This protocol permits the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
Detection of Total target protein with AlphaLISA SureFire Ultra reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol allows for miniaturization while maintaining AlphaLISA SureFire Ultra quality.
| Application |
Cell Signaling
|
|---|---|
| Automation Compatible |
Yes
|
| Brand |
AlphaLISA SureFire Ultra
|
| Detection Modality |
Alpha
|
| Lysis Buffer Compatibility |
Lysis Buffer
|
| Molecular Modification |
Total
|
| Product Group |
Kit
|
| Sample Volume |
10 µL
|
| Shipping Conditions |
Shipped in Blue Ice
|
| Target |
EGF Receptor
|
| Target Class |
Phosphoproteins
|
| Target Species |
Human
|
| Technology |
Alpha
|
| Therapeutic Area |
Immuno-oncology
Oncology
|
| Unit Size |
500 assay points
|
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