SRC (Human) Recombinant Protein
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Specification
Product Description
Human SRC (226 a.a. - 536 a.a.) partial recombinant protein with an N-terminal GST tag and a C-terminal His tag expressed in Baculovirus-infected sf9 cells.
Host
Human
Theoretical MW (kDa)
60
Form
Liquid
Preparation Method
Baculovirus infected insect cell (Sf9) expression system
Purification
GSH agarose and Ni-NTA agarose chromatography
Purity
> 70% by SDS-PAGE
Activity
Assay activity of Src in a 500 uL reaction containing 20mM Hepes KOH (pH7.5), 4 mM MgCl2, 2 mM MnCl2, 1 mM DTT, 50 mM [gamma 32P ATP (1 uCi), and 4ug of CycLex-"Tyrosine kinase-binding module". Start the reaction by adding 10 uL of the enzyme, diluted 10-fold in a buffer containing 20 mM Hepes KOH (pH7.5), 1 mM DTT, 0.03% Brij35. Incubate for 69 minutes at 30°C. Terminate the reaction by adding 600 uL of cold 10% TCA solution containing 0.2% Sodium pyrophosphate and stand on ice for 15 min. Filtrate acid insoluble material through GFC filters (Whatman Inc.), wash 4 times with 1% TCA and rinse filters with ethanol. Dry filters and count in a liquid scintillation counter.
Storage Buffer
In 20 mM Hepes-KOH, 50 mM NaCl, pH7.5 (1% BSA, 1 mM EDTA, 2 mM DTT, 0.03% Brij35 and 50 % glycerol)
Storage Instruction
Stable for 12 months at -70°C.
Avoid repeated freezing and thawing.Note
SRC (Human) Recombinant Protein (Cat# P6990) is a positive control designed to use for c-Src Kinase (Human) Assay/Inhibitor Screening Assay Kit (Cat# KA0058). The SRC (Human) Recombinant Protein should be added to the well at 1 unit/well. For instance, diluted this positive control 1:10, use 10 uL for 1 assay.
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Applications
SDS-PAGE
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