Pull-down assayUbiquitinUbiquitin-activating enzymeUbiquitin-binding proteinUbiquitin-conjugating enzymeUbiquitin-conjugation reactionUbiquitylation is a posttranslational modification that utilizes protein-protein binding interactions to regulate cellular processes. In ubiquitin signaling, a vast array of mono- and...
Pull-down assay Binding between an MBP tagged linearly fused dimer of the RING domain of RNF4 and the Ubc13~Ub conjugate was performed as described previously7. Production of the linearly fused dimer of the RING domain of RNF4 was described previously3,7 and was used in the pull-down assay...
4 Pull-down and proteomic analysis of HeLa cells using sAB-K29. a, Pull-down assay of HeLa cell lysate using sAB-K29. The bound proteins were separated on an SDS-PAGE gel. Gel slices were subjected to label-free quantitative mass spectrometry to identify the enriched proteins. Three ...
In order to detect robust levels of cleavage products, the assay was performed in the presence of 2 μM MG132. (F) Pull-down assay using cells treated with tunicamycin (15 μg/ml for 4 hr) was performed as in (C). Coexpression of gp78-C356G/H362A (gp78 mt) reverted increased ...
, 200 mM NaCl, 1 mM DTT) containing 200 µg/mL lysozyme with sonication. The supernatant was loaded on amylose resin (New England Biolabs) for 2 h at 4 °C with rotation. The beads were washed with lysis buffer. Beads containing proteins were used for MBP-pull down assay...
Recombinant GST-OGT proteins were used in pull-down assays, and GST-OGT pulled-down Cdc20 (Fig. 1C). We then assessed the association between overproduced Cdc20 and endogenous OGT (Fig. 1, D and E). Flag-Cdc20 coIPs with endogenous OGT, and the interaction becomes more robust after ...
Here, pull-down assay and LC-MS/MS analysis revealed that ubiquitin-specif... A Shiba,Y Kim,M Noguchi - 《Journal of Thoracic Oncology》 被引量: 3发表: 2017年 Ubiquitin-specific peptidase 5 facilitates cancer stem cell-like properties in lung cancer by deubiquitinating beta-catenin Background...
Neither wild-type nor mutant rbovISG15 was able to interact with the ISG15 E1 initiating enzyme, UBE1L, in an in vitro pull-down assay. Ovine (ov) ISG15 has three additional amino acids within the hinge region that were hypothesized to increase stability and the degree of interaction with...
Ubiquitinated p53, auto-ubiquitinated LIN41 and pull-down efficiency were monitored by immunoblotting. (g) Mono- and poly-ubiquitinated proteins were purified using α-Ub-FK2- agarose beads capable of detecting all linkage types. iLin41 cells were prepared as for TUBE assay but without p53 ...
. We also showed that endogenous N-Myc binds to endogenous Huwe1 in IMR32 neuroblastoma cells (Fig. 1b). Using purified GST–N-Myc in a pull-down assay, we confirmed that N-Myc interacts specifically with Huwe1in vitro(Fig. 1c). Next, we compared the affinity of the interaction ...