Protein samples were denatured in NuPAGE sample buffer (Invitrogen) supplemented with 5% (v/v) 2-mercaptoethanol and by subsequent heating to 95°C for 5 min. Denatured proteins were resolved on 12% (w/v) NuPAGE polyacrylamide gels run with MOPS buffer (Invitrogen). See Blue Plus, protein molecular weight markers (Invitrogen) were run on all NuPAGE gels. Instant Blue stain (Expedeon Ltd.) was used to visualize the protein bands. Western blotting to detect either TMV coat protein with an anti-TMV antibody, (AS-72203-2ML, Austral Biologicals, San Ramon, CA, United States) or the six C-terminal histidine residues with a monoclonal antibody prepared to six histidine residues, (Cat 34660, Qiagen) was performed as described by Saunders and Lomonossoff (2015) (link).
See blue plus
Manufactured by Thermo Fisher Scientific
See Blue Plus is a laboratory reagent designed to detect and visualize nucleic acids in gel electrophoresis. It is a fluorescent dye that binds to DNA or RNA, allowing for the identification and quantification of nucleic acid samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using see blue plus
1
SDS-PAGE and Western Blot Analysis
2
SDS-PAGE Protein Separation Protocol
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The analysis
was performed using Bolt 4–12% Bis-Tris Plus gels (10 wells)
at 165 V for 36 min utilizing prestained Invitrogen SeeBlue Plus2
molecular weight standard. Bolt LDS sample buffer (4×) was added
to each protein sample (1 μg) for loading on the gel. The bands
were visualized by Coomassie blue staining.
was performed using Bolt 4–12% Bis-Tris Plus gels (10 wells)
at 165 V for 36 min utilizing prestained Invitrogen SeeBlue Plus2
molecular weight standard. Bolt LDS sample buffer (4×) was added
to each protein sample (1 μg) for loading on the gel. The bands
were visualized by Coomassie blue staining.
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