CAMK2β, His-tag Recombinant

Catalog #

40041

$410 *

Size: 10 µg

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Description

Recombinant human CAMK2β (Calcium/calmodulin Dependent Protein Kinase II Beta), encompassing amino acids 1-503. This construct contains an N-terminal His-tag. The recombinant protein was affinity purified and is active.

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Synonyms

CAMKB, CAM2, CAMK2, MGC29528, CAMK-2

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Construct

CAMK2β (His-Full Length)

Host Species/Expression System

Sf9 insect cells

Purity

≥70%

Format

Aqueous buffer solution

Formulation

50 mM Sodium Phosphate, pH 7.0, 300 mM NaCl, 150 mM Imidazole, 0.1 mM PMSF, 0.2 mM DTT, 25% glycerol

58 kDa

Amino Acids

full length

Genbank #

NM_172081

UniProt #

Q13554

Tag(s)

N-terminal His-tag

Background

CAMK2β belongs to the serine/threonine protein kinase family and to the type II multifunctional Ca(2+)/calmodulin-dependent protein kinase subfamily. CAMK2β showed wide tissue and cell distribution, and one of CAMK2β variant predominated in adult brain (1). The ratio of CAMK2α and CAMK2β protein levels were inversely related during activity in hippocampal neurons (2). CAMK2β is a prominent kinase in the central nervous system and may function in long-term potentiation and neurotransmitter release.

References

1. Tombes, R. M. et al: Identification of novel human tumor cell-specific CaMK-II variants. Biochim. Biophys. Acta 1355: 281-292, 1997.
2. Thiagarajan, T. C. Et al: Alpha- and beta-CaMKII: inverse regulation by neuronal activity and opposing effects on synaptic strength. Neuron 36: 1103-1114, 2002.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

CAMK2β activity was measured by using the autocamtide 2 synthetic peptide substrate (KKALRRQETVDAL-amide), diluted in distilled water to a final concentration of 1 mg/ml, in the ADP Glo™ assay (Promega #V9101). Reaction was initiated by mixing increasing amounts of the CAMK2β with 25 µM ATP in 40 mM Tris-HCl, pH 7.4, 20 mM MgCl₂, 0.1 mg/ml BSA prepared with 250 µM DTT, co-factor calcium/calmodulin and substrate at a final concentration of 200 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 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

Kinase/Serine-Threonine

Regulatory Status

Research Use Only

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Datasheets/Protocols

40041 Lot 230419

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