The modification of proteins with ubiquitin is an important cellular mechanism for targeting abnormal or short-lived proteins for degradation. Ubiquitination involves at least three classes of enzymes: ubiquitin-activating enzymes, or E1s, ubiquitin-conjugating enzymes, or E2s, and ubiquitin-protein ligases, or E3s. This gene encodes a member of the E2 ubiquitin-conjugating enzyme family. This enzyme is required for post-replicative DNA damage repair. Its protein sequence is 100% identical to the mouse, rat, and rabbit homologs, which indicates that this enzyme is highly conserved in eukaryotic evolution.
Background References
1. Polge C. et. al. UBE2B is implicated in myofibrillar protein loss in catabolic C2C12 myotubes. Version 2. J Cachexia Sarcopenia Muscle. 2016 Jun
2. Mou L. et. al. A functional variant in the UBE2B gene promoter is associated with idiopathic azoospermia. Reprod Biol Endocrinol. 2015 Jul
Western blot analysis of Ube2B on different lysates with Rabbit anti-Ube2B antibody (ET7111-09) at 1/5,000 dilution.
Lane 1: HepG2 cell lysate (10 µg/Lane) Lane 2: Jurkat cell lysate (10 µg/Lane) Lane 3: NIH/3T3 cell lysate (10 µg/Lane) Lane 4: PC-12 cell lysate (10 µg/Lane) Lane 5: Mouse testis tissue lysate (20 µg/Lane) Lane 6: Rat testis tissue lysate (20 µg/Lane)
Predicted band size: 17 kDa Observed band size: 17 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7111-09) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Western blot analysis of Ube2B on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7111-09, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Ube2B antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7111-09, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human fetal skeletal muscle tissue using anti-Ube2B antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7111-09, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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