Recombinant PRMT5 / MEP50 complex

Catalog No: 31521 Format: 20 µg ¥4,040 Buy
Catalog No: 31921 Format: 1 mg ¥46,760 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

PRMT5 (Protein Arginine Methyltransferase 5, also known as JBP1, Janus Binding Protein 1) is a type II arginine methyltransferase, a protein that transfers methyl groups to the arginine residues of substrate proteins. Arginine methylation is a common post-translational modification of histones and other cellular proteins. PRMT5 symmetrically dimethylates histone H2A Arg3, H3 Arg8 and H4 Arg3. It also methylates Piwi proteins which regulate small non-coding RNAs and snRNPs, regulating snRNP core particle assembly. PRMT5 functions as a co-repressor of transcription and is recruited by a number of DNA binding proteins and transcriptional repressor complexes. PRMT5 acts as part of a multimeric complex in concert with a variety of partner proteins that regulate its function and specificity. A core component of these complexes is the WD40 protein MEP50/WDR77/p44, which mediates interactions with binding partners and substrates. PRMT5 is present both in the nucleus and the cytoplasm. It also interacts with COPR5, JAK2, PRDM1 and RPS10.

Application Notes

Recombinant PRMT5 / MEP50 Complex is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

HMT Assay Conditions: 0.5 µg Histone H4 (Cat. No. 31493) was incubated with different concentrations of PRMT5 / MEP50 Complex in 30 µl reaction system including 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, and 50 µM SAM for 3 hours at room temperature. Activity was detected by Western Blot.

The following applications have been published using this product. Unless noted above, Active Motif may not have validated this recombinant protein for use in these applications:

Methylation Assay

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Protein Details

Recombinant PRMT5 / MEP50 Complex that includes full-length PRMT5 (accession numbers NP_006100.2) with an N-terminal FLAG tag and full-length MEP50 (accession numbers NP_077007.1) without tag was expressed in a baculovirus expression system. The molecular weights of PRMT5 and MEP50 are 74 kDa and 36.7 kDa, respectively.

 

Recombinant PRMT5 / MEP50 complex - protein gel.
10% SDS-PAGE gel stained with Coomassie blue.
MW of PRMT5: 74 kDa
MW of MEP50: 36.7 kDa
Purity: > 90%

PRMT / MEP50 complex, Activity Assay
0.5 µg Histone H4 (Cat# 31493) was incubated with 0 µg (-), 0.225 µg (+), 0.45 µg (++), or 0.9 µg (+++) PRMT5 / MEP50 complex in 30 µl reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 50 µM SAM for 3 hours at room temperature. 6 µl reaction products were loaded and run on a 12.5% SDS-PAGE gel. Western Blot was used to detect the generation of reaction products (H4R3me2s antibody, Cat. No. 61187, 1:1000 dilution). PRMT5 / MEP50 Complex only was used as negative control.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.