Optimization and application of silver staining of non-glycosylated and glycosylated proteins and nucleic acids for agarose native gel electrophoresis
| Autor: | Teruo Akuta, Takashi Shibata, Masataka Nakagawa, Chiaki Sakuma, Yasuo Ono, Yasunori Kurosawa, Yoshinori Sato, Ryo Sato, Yui Tomioka, Tsutomu Arakawa |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Silver Staining
Biochemistry Stain Silver stain chemistry.chemical_compound Structural Biology Nucleic Acids Animals Humans Molecular Biology Electrophoresis Agar Gel chemistry.chemical_classification SARS-CoV-2 Silver Staining Method Egg Proteins Orosomucoid General Medicine Staining Isoelectric point chemistry Spike Glycoprotein Coronavirus Nucleic acid Agarose Glycoprotein Chickens |
| Zdroj: | International Journal of Biological Macromolecules. 189:869-878 |
| ISSN: | 0141-8130 |
| DOI: | 10.1016/j.ijbiomac.2021.08.142 |
| Popis: | Electrophoresis is one of the major techniques to analyze macromolecular structure and interaction. Its capability depends on the sensitivity and specificity of the staining methods. We have here examined silver staining of proteins and nucleic acids separated by agarose native gel electrophoresis. By comparing five commercial kits, we identified Silver Stain Plus from Bio-Rad most adequate, as it provided little background staining and reasonable band staining. One of the disadvantages of the Silver Stain Plus kit is its variable staining of glycoproteins as tested with several model samples, including hen egg white proteins, α1-acid glycoprotein and SARS-CoV-2 Spike protein. One of the advantages of silver staining is its ability to stain nucleic acids as demonstrated here for a model nucleic acid with two kits. It was then used to monitor the removal of nucleic acids from the affinity-purified maltose binding protein and monoclonal antibody. It also worked well on staining proteins on agarose gels prepared in the vertical mode, although preparation of the vertical agarose gels required technological modifications described in this report. With the silver staining method optimized here, it should be possible in the future to analyze biological samples that may be available in limited quantity. |
| Databáze: | OpenAIRE |
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