IndraLab

Statements

GST binds USP14. 10 / 10
| 1 9
sparser
"We purified USP14 and USP14(C114A) and their glutathione S-transferase (GST)-conjugated counterparts [GST-USP14 and GST-USP14(C114A), respectively] using previously described methods7, with some modifications."
26041011
sparser
"To identify USP14 aptamers, we used SELEX techniques with GST-USP14 (78 kDa), while USP14 (53 kDa) was also expressed and purified for subsequent analysis with aptamers and proteasomes (Fig. 1A)."
26041011
sparser
"The random RNA library contained random 40-nt sequences, flanked by a 3' region (23 bp) and a sequence containing the T7 promoter (46 bp, underlined). (C) Scheme for the SELEX strategy. (1) Purified RNAs were incubated with GST-USP14. (2) USP14-RNA complexes were captured by glutathione agarose beads. (3) Unbound RNA molecules were removed by centrifugation."
26041011
reach
"Endogenous Usp14 protein derived from mouse brain lysates specifically bound to the immobilized GST tagged GABA A R alpha1 loop, but not to GST alone (XREF_FIG), indicating in vitro binding of the protease and the receptor polypeptide."
19759851
sparser
"Purified GST (data not shown), GST-USP14, GST-USP14(C114A), USP14, and USP14(C114A) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and gels were stained with Coomassie Brilliant Blue R-250 (CBB) to determine the size and purity of proteins (Fig. 1A)."
26041011
sparser
"The random RNA library contained random 40-nt sequences, flanked by a 3’ region (23 bp) and a sequence containing the T7 promoter (46 bp, underlined). (C) Scheme for the SELEX strategy. (1) Purified RNAs were incubated with GST-USP14. (2) USP14-RNA complexes were captured by glutathione agarose beads. (3) Unbound RNA molecules were removed by centrifugation. (4) USP14-RNA complexes were dissociated with elution buffer containing excess imidazole. (5) RNAs bound to USP14 were prepared by phenol:chloroform extraction and ethanol precipitation."
26041011
sparser
"We loaded and immobilized 2 pmol of GST-USP14 or GST onto a GST sepharose 4B resin, and then added 20 pmol of RNA aptamers."
26041011
sparser
"Purification of USP14 and human proteasomes, preparation of vme-proteasomes, and SELEX for USP14 aptamers. (A) Approximately 1 μ g of purified recombinant USP14, USP14(C114A), GST-USP14, and GST-USP14(C114A) was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining."
26041011
sparser
"After washing with PBS, GST-USP14 was eluted using 10 mM reduced glutathione (50 mM Tris-HCl pH 8.0)."
26041011
sparser
"Figure 1: (A) Approximately 1 μg of purified recombinant USP14, USP14(C114A), GST-USP14, and GST-USP14(C114A) was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining."
26041011