Western blot analysis of extracts from various cell lines using Ubc9 Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. 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 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 June 2020
Protocol Id: 10
Ubc9 Antibody detects endogenous levels of total Ubc9 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding V148 of human Ubc9. Antibodies are purified by peptide affinity chromatography.
The process of SUMO-1 conjugation is similar to that seen with ubiquitin and other forms of post-translational protein modification (1). Like ubiquitin, SUMO-1 is conjugated to its target protein by the coordinated action of ubiquitin conjugation enzymes E1, E2 and E3 (2). Ubc9 (or ube2M) is a highly conserved, 158 amino acid protein that acts as a SUMO-1 conjugating enzyme (3). Ubc9 binds to target proteins through their SUMO-1-CS (consensus sequence) domains and interacts with SUMO via the structurally conserved amino-terminal domain (3,4). Localization of Ubc9 to the nucleus and the nuclear envelope allows this enzyme to catalyze target protein sumoylation and regulate target protein nucleocytoplasmic transport and transcriptional activity (5,6). Ubc9 target proteins include a host of proteins (RAD51, RAD52, p53 and c-Jun) that regulate the cell cycle, DNA repair, and p53-dependent processes (7).
Explore pathways + proteins related to this product.