FGFR1 (V561M), GST-tag Recombinant

Catalog #

40209

$410 *

Size: 10 µg

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Description

Recombinant human FGFR1 (fibroblast growth factor receptor 1), encompassing amino acids 399-822. This construct contains an N-terminal GST-tag and the mutation of interest V561M. The recombinant protein was affinity purified and is active.

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Synonyms

CEK; FLG; FLT2; KAL2; BFGFR; CD331; FGFBR; HBGFR; N-SAM; C-FGR, FGFR-1

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

FGFR1 (V561M) (GST-399-822)

Mutation

V561M

Host Species/Expression System

Sf9 insect cells

Purity

≥90%

Format

Aqueous buffer solution

Formulation

50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM Glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF, and 25% glycerol

75 kDa

Amino Acids

399–822

Specific Activity

30 pmol/min/µg

Genbank #

NM_023110

UniProt #

P11362

Tag(s)

N-terminal GST-tag

Background

FGFR1 (also known as FLT2) is a member of the Fibroblast Growth Factor Receptor family that constitute a family of four membrane-spanning tyrosine kinases (FGFR1-4) which serve as high-affinity receptors for 17 growth factors (FGF1-17). The FGF Receptor family plays an important role in multiple biological processes, including mesoderm induction and patterning, cell growth and migration, organ formation and bone growth (1). FGFR1 is alternatively spliced generating multiple splice variants that are differentially expressed during embryo development and in the adult body (2).

References

1. Xu, X. et al: Fibroblast growth factor receptors (FGFRs) and their roles in limb development. Cell Tissue Res. 1999 Apr;296(1):33-43.
2. Groth, C. et al: The structure and function of vertebrate fibroblast growth factor receptor 1. Int J Dev Biol. 2002;46(4):393-400.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

FGFR1 (V561M) activity was measured by using the Poly(Glut4Tyr1) synthetic peptide, diluted in 25 mM Tris-HCl (pH 7.5) to a working concentration of 1 mg/ml, in the ADP Glo™ Kinase Assay kit (Promega #V9101). Reaction was initiated by mixing increasing amounts of FGFR1 (V561M) with 25 μM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MnCl₂, 0.1 mg/ml BSA prepared with 50 μM DTT, 12.5 mM MgCl₂ and substrate at a final concentration of 200 µg/ml.

After a 40-minute incubation at room temperature, the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at room temperature. Kinase Detection Reagent was added, and the reaction was incubated for another 30 minutes at ambient temperature. Detection of luminescence was measured using the Luminescence Module Protocol on GloMax®-Multi Microplate Multimode Reader. The Specific Activity was calculated as follows: (Corrected activity, RLU) / [(Specific activity from ADP in RLU/pmol) * (Reaction time in min) *(Enzyme amount in μg or mg)]. Corrected RLU was calculated by subtracting the blank value from all the values. The blank was determined from a “no enzyme” sample by replacing the enzyme 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

Warnings

Avoid freeze/thaw cycles

Scientific Category

Tyrosine Kinase

Regulatory Status

Research Use Only

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Datasheets/Protocols

40209 Lot 240126

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