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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. It is present both in the nucleus and the cytoplasm. It also interacts with COPR5, JAK2, PRDM1 and RPS10.
Application Notes
Recombinant PRMT5 is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. PRMT5 is an active H3R8 and H4R3 methyltransferase.
HMT Assay Conditions: 0.5 µg Histone H4 (Cat. No. 31493) was incubated with different concentrations of PRMT5 protein in 30 µl reaction system including 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. Activity was detected by Western Blot.
Protein Details
Recombinant PRMT5 (accession number NP_006100.2) was expressed in Sf9 cells and contains an N-terminal FLAG tag with an observed molecular weight of 74.2 kDa. The recombinant protein is >70% pure by SDS-PAGE.
References
The following applications have been published using this protein. Unless noted above, Active Motif may not have validated this product for use in these applications:
Methylation Assay
View publications that use Active Motif products & services here. Enter the product number(s) to see publications & applications that use this product.
Recombinant PRMT5 protein gel.
PRMT5 protein was run on a 10% SDS-PAGE gel and stained with Coomassie Blue.
MW: 74 kDa
Purity: > 90%
Recombinant PRMT5 activity assay.
0.5 µg of Histone H4 (Cat. No. 31493) was incubated with 0 µg (-), 0.15 µg (+), 0.3 µg (++), and 0.6 µg (+++) PRMT in 30 µl reaction volume containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 50 uM SAM for 3 hr at room temp. 6 µl of reaction products were run on a 12.5% SDS-PAGE and detected by Western Blot using H4R3me2s antibody (Cat# 61187) at 1:1000. PRMT5 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 guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
