The turnover of ΔssCPY∗-GFP and Hmg2-myc was assessed in cells by inhibiting protein synthesis with cycloheximide. Cells were collected at the indicated time points and ΔssCPY∗-GFP and Hmg2-myc levels analyzed by immunoblotting and quantified by densitometry. Values shown are percentages relative to zero time points from three independent biological repeats. Protein extracts were electrophoresed under reducing conditions on NuPAGE minigels (Thermo Fisher Scientific) and electroblotted onto polyvinylidene fluoride membrane (GE Healthcare). Primary antibodies used were rabbit α-Sup35 (Ness et al., 2002 (link)), ubiquitin (sc-8017; Santa Cruz Biotechnology, Inc.), Ydj1 (ab74442; Abcam), Sis1 (COP-080051; Operon Biotechnologies), Ssa1 (ADI-SPA-822; Enzo Life Sciences), GFP (A6465, Invitrogen), Pgk1 (459250; Thermo Fisher Scientific), Hsp104 (ab2924; Abcam), rabbit α-Sse1 (Chiabudini et al., 2012 (link)), and Myc 4A6 (05–724; EMD Millipore).
Ab2924
Manufactured by Abcam
Ab2924 is a laboratory equipment product. It is a laboratory instrument designed for a specific functional purpose. No further details are available without the risk of bias or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Ubiquitin sc 8017, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene fluoride membrane, by GE Healthcare (1 mentions)
Nupage minigels, by Thermo Fisher Scientific (1 mentions)
Myc 4a6, by Merck Group (1 mentions)
Pvdf membrane, by Cytiva (1 mentions)
Hsp104, by Abcam (1 mentions)
Odyssey fc, by LI COR (1 mentions)
2 protocols using ab2924
1
Protein Turnover Dynamics Analysis
2
Protein Immunoblotting Analysis
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Protein extracts were electrophoresed under reducing conditions on SDS-PAGE minigels and electroblotted onto PVDF membrane (Amersham Pharmacia Biotech). Bound antibody was visualized using LI-COR Odyssey-FC. Primary antibodies used were Pgk1 (459250, ThermoFisher Scientific) and Hsp104 (ab2924, Abcam).
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