EGFR (V948R), His-Tag Recombinant

Catalog #

101644

$410 *

Size: 10 µg

Qty

Bulk/Custom

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Description

Recombinant human EGFR (epidermal growth factor receptor), encompassing amino acids (699-1022) length. This construct contains an N-terminal His-tag and mutation of interest (V948R). The recombinant protein was affinity purified and is kinase active.

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Synonyms

ERBB, mENA, ERBB1, HER1

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

EGFR (V948R) (His-669-1022)

Mutation

V948R

Host Species/Expression System

Sf9

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM Sodium Phosphate, pH 7.0, 300 mM NaCl, 150 mM Imidazole, 0.25 mM DTT, 25% glycerol

48 kDa

Amino Acids

669-1022

Specific Activity

38 pmol/min/µg

Genbank #

NM_005228

UniProt #

P00533

Tag(s)

N-terminal His-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

EGFR (V948R) activity was measured by using Poly (4:1 Glu, Tyr) synthetic peptide substrate diluted in 25 mM Tris-HCl buffer (pH 7.5) to a final concentration of 1 mg/ml in the ADP Glo™ assay (Promega #V9101). Reaction was initiated by mixing increasing amounts of the EGFR (V948R) with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 250 µM DTT and substrate with a final concentration of 50 µg/ml.

After a 40-minute incubation at 37°C, the reaction was terminated by addition of the AMP-Glo™ Reagent followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was then added and incubated for another 30 minutes. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate reader. The corrected activity (RLU) was calculated by removing the blank value for each sample divided by the (specific activity of ADP in RLU/pmol)*(Reaction time in min)*(Enzyme amount in µg or mg). The blank was determined from a “no kinase” sample by replacing the enzyme working solution with an equal volume of Kinase Dilution Buffer X (1X).

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Scientific Category

Tyrosine Kinase

Regulatory Status

Research Use Only