ROS1 (G2101A), GST-Tag Recombinant

Catalog #

101641

$410 *

Size: 10 µg

Qty

Bulk/Custom

Add to Favorites

*US Pricing only. For international pricing, please contact your local distributor.

●

Purchase

●

Description

Recombinant human ROS1 (G2101A) (ROS proto-oncogene 1, receptor tyrosine kinase), encompassing amino acids 1881-end. This construct contains an N-terminal GST-tag and was affinity purified.

●

Synonyms

Proto-oncogene tyrosine-protein kinase ROS, MCF3, ROS

●

Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

ROS1 (G2101A) (GST-1881-end)

Mutation

G2101A

Host Species/Expression System

Sf9

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 300 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, 25% glycerol

82 kDa

Amino Acids

1881-end

Specific Activity

131 pmol/min/µg

Genbank #

NM_002944

UniProt #

P08922

Tag(s)

N-terminal GST-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

Kinase activity was measured using the ADP-Glo™ Kinase Assay Kit (Promega; Cat# V9101) which quantifies the amount of ADP produced. The ADP-Glo™ Reagent is added to terminate the reaction and deplete the remaining ATP. The Kinase Detection Reagent is then added to convert ADP to ATP and to measure the newly synthesized ATP using a luciferase reaction.

ROS1 activity was measured by using IGFR1tide synthetic peptide (KKKSPGEYVNIEFG) diluted in water to a final concentration of 1 mg/ml. Reaction was initiated by mixing increasing amounts of the ROS1 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 IX (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