Western blot analysis of extracts from various cell lines using PRMT7 (D1K6R) Rabbit mAb.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
PRMT7 (D1K6R) Rabbit mAb recognizes endogenous levels of total PRMT7 protein.Species Reactivity:
Human, Mouse, MonkeySpecies predicted to react based on 100% sequence homology:
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val233 of human PRMT7 protein.
Protein arginine N-methyltransferase 7 (PRMT7) is a member of the protein arginine N-methyltransferase (PRMT) family of proteins that catalyze the transfer of a methyl group from S-adenosylmethionine (AdoMet) to a guanidine nitrogen of arginine (1). The three types of PRMTs share the ability to mono-methylate arginine residues, but vary in their ability to generate differential methylation states (1-3). Mono-methylated arginine residues are further methylated by type I PRMTs to generate an asymmetric di-methyl arginine or by type II PRMTs to form a symmetric-dimethyl arginine. Type III methyltransferases are only able to mono-methylate arginine residues (1-3). Research studies indicate that PRMT7 is a type III PRMT that displays substrate specificity for an arginine-X-arginine (RXR) motif surrounded by several basic residues (4,5). PRMT7 interacts with a wide array of protein substrates and likely plays a role in many biological processes including pluripotency, neuronal differentiation, genomic instability, snRNP biogenesis, and breast cancer metastasis (6-11).
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