MEKK3, GST-tag Recombinant

Catalog #

40124

$410 *

Size: 10 µg

Qty

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Description

Recombinant human MEKK3 (MAPK/ERK kinase kinase 3, also known as MAP3K3), full length. This construct contains an N-terminal GST-tag. The recombinant protein was affinity purified.

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Synonyms

Mitogen-activated protein kinase kinase kinase 3, MAPK/ERK kinase kinase 3, MEK kinase 3, MEKK 3, MAP3K3, MAPKKK3

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

MEKK3 (GST-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥75%

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

105 kDa

Amino Acids

full length

Specific Activity

23 pmol/min/µg

Genbank #

NM_002401

UniProt #

Q99759

Tag(s)

N-terminal GST-tag

Background

MEKK3 is a member of the MEKK family of protein kinases that directly regulate SAPK and ERK pathway but not the p38 pathway. MEKK3 modulates SAPK and ERK activity by activating the upstream targets SEK and MEK1/2, respectively (1). KSR2 can regulate the activity of MEKK3 and can reduce MEKK3-mediated ERK activation. Knockout of the MEKK3 gene in mice leads to disruption of blood vessel development and the structural integrity of the yolk sac (2). The blockage of angiogenesis in these mice is not a result of decreased VEGF1 expression but rather due to an intrinsic defect in Mekk3 -/- endothelial cells. Thus, MEKK3 is necessary for blood vessel development and may be a possible target for drugs that control angiogenesis.

References

Ellinger-Ziegelbauer, H. Et al: Direct activation of the stress-activated protein kinase (SAPK) and extracellular signal-regulated protein kinase (ERK) pathways by an inducible mitogen-activated protein kinase/ERK kinase kinase 3 (MEKK) derivative. J. Biol. Chem. 272: 2668-2674, 1997.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

MEKK3 kinase activity was measured using MBP (Myelin Basic Protein) as substrate. Increasing amounts of the protein kinase were mixed with a final concentration of 0.25 mg/ml MBP in a buffer consisting of 5 mM MOPS pH 7.2, 2.5 mM β-glycerol-phosphate, 5 mM MgCl2, 1 mM EGTA, 0.4 mM EDTA, 25 µg/ml BSA (bovine serum albumin), and 100 µM DTT (final concentrations).
The reaction was initiated by adding [33P]-ATP (1 µCi/sample) mixed with non-radioactive ATP (final concentration of 50 µM). The reaction was incubated for 15min at 30°C and terminated by spotting 20 μl of the mixture onto phosphocellulose paper strips that were fixed in 1% phosphoric acid and washed three times. Radioactivity was determined using a scintillation counter. The blank was determined from a “no substrate” sample.

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/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40124 Lot 220210

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