MKK4, His-Tag Recombinant

Catalog #

101874

$410 *

Size: 10 µg

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Description

Recombinant human full-length MKK4 (dual-specificity mitogen-activated protein kinase kinase 4, also known as MAP2K4 or MAPKK4). This construct contains an N-terminal His-tag. The recombinant protein was affinity purified and is active.

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Synonyms

Dual specificity mitogen-activated protein kinase kinase 4, MAP2K4, JNKK1, MEK4, MP2K4, PRKMK4, SEK1, SERK1, SKK1

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

MKK4 (His-Full Length)

Host Species/Expression System

Sf9

Purity

≥70%

Format

Aqueous buffer solution

Formulation

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

46 kDa

Amino Acids

full length

Specific Activity

109 pmol/min/µg

Genbank #

NM_003010

UniProt #

P45985

Tag(s)

N-terminal His-tag

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

MKK4 activity was measured by using unactive 0.1 mg/ml JNK1 protein, diluted in 50 mM Tris-HCl, pH 7.5, 50 mM NaCl, 10 mM glutathione, 0.1 mM EDTA, 0.25 mM DTT, 0.1 mM PMSF and 25% glycerol, in the ADP Glo™ kinase assay (Promega #V9101). Reaction was initiated by mixing increasing amounts of the MKK4 with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 25 mM MgCl₂, 0.1 mg/ml BSA prepared with 50 µM DTT and substrate at a final concentration of 20 µg/ml final concentration.

After a 40-minute incubation at Room Temperature (RT), the reaction was terminated by addition of ADP-Glo™ Reagent, followed by a subsequent 40-minute incubation at RT. 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 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

Warnings

Avoid freeze/thaw cycles

Scientific Category

Dual Kinase

Regulatory Status

Research Use Only

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Datasheets/Protocols

101874 Lot 230608

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