Recombinant human PRMT5

Catalog # : EPX-012-RBV

Source : Human

Expressed in : SF9 cells

Quantity : 10µg of enzymatically active recombinant PRMT5 at 0.5µg /µl

Background:

Arginine methyltransferase that can both catalyze the formation of omega-N monomethylarginine (MMA) and symmetrical dimethylarginine (sDMA), with a preference for the formation of MMA (1). Methylates histone H2A and H4 'Arg-3' during germ cell development. Methylates histone H3 'Arg-8', which may repress transcription (2).

Protein details:

Recombinant human N-terminal Flag tagged PRMT5 was produced in SF9 insect cells, purified using FPLC and formulated in a storage buffer containing 20mM Tris-Cl pH 7.6, 1mM EDTA, 0.15 M NaCl, 10% glycerol, 0.5mM PMSF and 1mM DTT. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

Quality control:

Each lot has been evaluated by 10% Tris Glycine SDS-PAGE.

SDS-PAGE gel of recombinant Flag tagged  PRMT5 (Lane 2).  Lane 1, protein molecular weight marker.

HAT assay:

Recombinant human histone H3-H4 tetramers (2 μg) were acetylated in a 25-μl reaction containing 50 mM Tris HCl, pH 8.0, 10% glycerol, 0.1 mM EDTA, 1 mM DTT, 3 nM at 5 nCi/μl of [3H] acetyl-CoA (Amersham Pharmacia), and 0.2 pmol of purified recombinant FLAG-PRMT5. The mixture was incubated at 30°C for 1 h.

Storage:

-80°C

Guarantee:

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

Purity:

>98% purity by SDS PAGE.

Sequence:

PRMT5 (Mw: 72,202 Da.)
MDYKDDDDKRGPNSGTEKGRLVIPEKQGFDFLCMPVFHPRFKREF
IQEPAKNRPGPQTRSDLLLSGRDWNTLIVGKLSPWIRPDSKVEKI
RRNSEAAMLQELNFGAYLGLPAFLLPLNQEDNTNLARVLTNHIHT
GHHSSMFWMRVPLVAPEDLRDDIIENAPTTHTEEYSGEEKTWMWW
HNFRTLCDYSKRIAVALEIGADLPSNHVIDRWLGEPIKAAILPTS
IFLTNKKGFPVLSKMHQRLIFRLKLEVQFIITGTNHHSEKEFCSY
LQYLEYLSQNRPPPNAYELFAKGYEDYLQSPLQPLMDNLESQTYE
VFEKDPIKYSQYQQAIYKCLLDRVPEEEKDTNVQVLMVLGAGRGP
LVNASLRAAKQADRRIKLYAVEKNPNAVVTLENWQFEEWGSQVTV
VSSDMREWVAPEKADIIVSELLGSFADNELSPECLDGAQHFLKDD
GVSIPGEYTSFLAPISSSKLYNEVRACREKDRDPEAQFEMPYVVR
LHNFHQLSAPQPCFTFSHPNRDPMIDNNRYCTLEFPVEVNTVLHG
FAGYFETVLYQDITLSIRPETHSPGMFSWFPILFPIKQPITVREG
QTICVRFWRCSNSKKVWYEWAVTAPVCSAIHNPTGRSYTIGL

References:

1.    Antonysamy S., Bonday Z., Campbell R.M., Doyle B., Druzina Z., Gheyi T., Han B., Jungheim L.N., Qian Y., Rauch C., Russell M., Sauder J.M., Wasserman S.R., Weichert K., Willard F.S., Zhang A., Emtage S.Proc. Natl. Acad. Sci. U.S.A. 109:17960-17965(2012)
2.    Pal S., Vishwanath S.N., Erdjument-Bromage H., Tempst P., Sif S. Mol. Cell. Biol. 24:9630-9645(2004)