RIPK3, GST-Tag Recombinant

Catalog #

79710

$410 *

Size: 10 µg

Qty

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Description

Recombinant human full-length RIPK3 (also known as receptor-interacting serine/threonine-protein kinase 3 or RIP3). This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified and is active.

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Synonyms

Receptor interacting serine/threonine kinase 3, RIP-like protein kinase 3, Receptor-interacting protein 3, RIP3, RIP-3

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

RIPK3 (GST-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥70%

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, 25% glycerol

96 kDa

Amino Acids

Full Length

Specific Activity

9.5 pmol/min/µg

Genbank #

NM_006871

UniProt #

Q9Y572

Tag(s)

N-terminal GST-Tag

References

1. Kasof, G. M. et al., FEBS Lett. 47: 285-291, 2000.
2. Oberst, A. et al., Nature. 471: 3636-367. 2011.

Storage/Stability

At least 6 months at –80°C.

Assay Conditions

RIPK3 activity was measured by using MBP protein, diluted in 100 mM MOPS to a final concentration of 0.5 mg/ml, in the ADP Glo™ assay (Promega #V9101). Reaction was initiated by mixing increasing amounts of the RIPK3 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 2.5 mM MgCl₂, 0.1 mg/ml BSA prepared with 50 µM DTT, 2.5 mM co-factor MnCl₂ and substrate at a final concentration of 500 µg/ml final concentration.
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 corrected activity (RLU) was calculated by removing the blank value for each sample. The Kinase Specific Activity was calculated as follows: RLU / [(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 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

Kinase

Regulatory Status

Research Use Only

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Datasheets/Protocols

79710 Lot 230525

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