Journal: Current protocols in protein science
Article Title: Protein Detection in Gels Using Fixation
doi: 10.1002/0471140864.ps1005s29
Figure Lengend Snippet: Representative Common Silver Stain Kits
Article Snippet: The protocol for SYPRO Ruby staining described below has been adapted from the method described by the manufacturer of the fluorophore (Thermo Fisher). table ft1 table-wrap mode="anchored" t5 caption a7 Representative Vendors Stain Limit of Detection Time MS Compatible BioRad Dodeca Silver Stain Kit Silver Stain Plus Kit Silver Stain 0.25–0.5 ng 0.6–1.2 ng 0.6–1.2 ng 3 hr 1.5 hr 2 hr yes yes Thermo Fisher Pierce Silver Stain for Mass Spec Pierce Silver Stain Kit Pierce Color Silver Stain Kit SilverXpress ® Silver Staining Kit 0.25 ng 0.25 ng 0.1 ng 0.86 ng 1 hr 13 min 2 hr 25 min 4 hr 5 min 2 hr yes yes yes yes Sigma ProteoSilver TM Plus Silver Stain Kit ProteoSilver TM Silver Stain Kit 0.1 ng 0.1 ng 1 hr 45 min 1 hr 45 min yes yes GE Healthcare PlusOne Silver Staining Kit, Protein 0.2–0.6 ng 2 hr yes a G Biosciences FASTsilver TM FOCUS TM FASTsilver TM 1 ng 1 ng 1 hr >90 min yes yes Amresco Silver Bullit TM Silver Stain Kit >1 ng 4 h 20 min yes Open in a separate window a This kit is compatible with mass spectrometry only when glutaraldehyde is omitted from the staining protocol Representative Common Silver Stain Kits Materials list-behavior=simple prefix-word= mark-type=none max-label-size=0 SDS 1- or 2-D polyacrylamide gels containing proteins of interest ( UNITS 10.1–10.4 ) Fixing solution: 40% (v/v) methanol/10% (v/v) acetic acid in H2O SYPRO Ruby protein gel stain (Thermo Fisher) Destain/wash solution: 2% (v/v) acetic acid 2% (w/v) glycerol Plastic container with tight-fitting lid, of size appropriate for gel Platform shaker Additional reagents and equipment for acquiring gel images (see Support Protocol) list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Remove polyacrylamide gel from the electrophoresis unit and place in a plastic container containing at least 20 gel volumes of 50% methanol/7% acetic acid.
Techniques: Silver Staining, Staining, Mass Spectrometry
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