Alternate Name: UBE2L3, ubiquitin-conjugating enzyme E2L 3, UBCH7 Domain Information: Ubiquitin-conjugating enzyme, Ubiquitin-conjugating enzyme E2, catalytic domain homologues Gene Ontology Terms: protein modification, ubiquitin--protein ligase, ubiquitin conjugating enzyme, ubiquitin-dependent protein degradation Application(s): Peptide ELISA: antibody detection limit dilution 1:128,000. Western Blot: Approx 18kD band observed in Hela and Jurkat lysates (predicted MW of 18kD according to NP_003338). Recommended for use at 0.5-2ug/ml. Staining (0.5ug/ml) of Hela lysate (RIPA buffer, 35ug total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Peptide Blocking: Corresponding peptide is available for Peptide Blocking studies. See U1000-88-P. Antibody is typically 0.5mg/ml and peptide is supplied as a 100ul pellet. When peptide is reconstituted in 200ul water, the concentration would also be 0.5mg/ml. To start, the best ratio would be 1:1 (molar excess of peptide relative to antibody when identical volumes are mixed). Mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. It is best is to have two identical blots to be incubated with equal amount of antibody, but one with the antibody pre-adsorbed to the peptide for 20min. Then incubate and develop the two blots in parallel.
