|H M R Mk||Endogenous||19||Rabbit IgG|
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
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.
UBE2G2 (D8Z4G) Rabbit mAb recognizes endogenous levels of total UBE2G2 protein. This antibody does not cross-react with UBE2G1 protein.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human UBE2G2 protein.
Ubiquitin (Ub) is a conserved polypeptide that is covalently linked to many cellular proteins through the process of ubiquitination, which targets proteins for degradation by the 26S proteasome. Three enzymatic components are involved in the protein ubiquitination cascade. Ubiquitin is first activated by forming a thioester complex with an E1 ubiquitin-activating enzyme. Activated ubiquitin is subsequently transferred to an E2 ubiquitin-carrier protein, and then from the E2 to an E3 ubiquitin ligase for final delivery to the ε-amino group of the target protein lysine residue (1-3).
The ubiquitin-conjugating enzyme E2 G2 (UBE2G2, UBC7) is a ubiquitously expressed E2 enzyme and critical component of the endoplasmic reticulum-associated degradation pathway (ERAD) (4). Research studies demonstrate that UBE2G2 forms homodimers and preassembles K48-linked poly-Ub chains at its active site (5-8). The association of Ub-charged UBE2G2 molecules with the ER-resident E3 ligase AMFR (gp78) is required for Ub chain transfer and efficient removal of misfolded or aggregated proteins through the ERAD pathway (9,10).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|63182S||100 µl (10 western blots)||$ 255.0|