title: A simple and cost-effective solid-phase protein nano-assay using polyacrylamide-coated glass plates.
abstract: A new solid-phase protein nano-assay is suggested for simple and sensitive estimation of protein content in sample buffers (a 1-μl sample is sufficient for analysis). The assay is different from conventional "on-filter" assays in that it uses inexpensive fully transparent polyacrylamide gel (PAAG)-coated glass plates as solid support and, thus, combines the convenience of "on-membrane" staining with the sensitivity and ease of documentation of "in-gel" staining (and, therefore, is especially suited for standard lab gel documentation systems). The PAAG plates assay is compatible with all dyes for in-gel protein staining. Depending on the sensitivity of the staining protocol, the assay can be used in macro-, micro-, and nano-assay formats. We also describe a low-cost two-component colloidal Coomassie brilliant blue G-250 (CBB G-250) staining protocol for fast quantitative visualization of proteins spotted on a PAAG plate (the detection limit is up to 2 ng of proteins even when using a Nikon CoolPix digital camera and white light transilluminator instead of a gel scanner). The suggested colloidal CBB G-250 protocol could also be used for visualizing nano-amounts of proteins in polyacrylamide gels. The PAAG plate assay could be useful for proteomic applications and, in general, for all cases where a fast, sensitive, and easily documentable cost-effective solid-phase protein assay is required.
authors.affiliation: Institute of Developmental Biology, Russian Academy of Sciences, Moscow 119334, Russia. Electronic address: wkrajewski@hotmail.com.
authors.forename: Wladyslaw A
authors.initials: WA
authors.lastname: Krajewski
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry*; D000818:Animals; D002021:Buffers; D002417:Cattle; D003362:Cost-Benefit Analysis; D005898:Glass / Q000737:chemistry*; D057230:Limit of Detection; D036103:Nanotechnology / Q000191:economics* / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis; D013997:Time Factors; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D002021:Buffers; D011506:Proteins; D012710:Serum Albumin, Bovine; D000099166:Polyacrylamides
keywords: Gel staining; Protein determination; Protein nano-assay; Solid-phase assay