title: Modified silver staining in 2DE improves protein detection even at extremely low sample concentration.
abstract: The typical concentration of protein loaded varies from 0.13 to 1.40 μg/μL for a classical silver staining method in 2DE gel. Here, we present a simple modified classical silver staining method by modifying the silver impregnation and development reaction steps. This modified method detects the protein spots at extremely low loaded concentrations, ranging from 0.0048 to 0.0480 μg/μL. We recommend this modified silver staining as an excellent method for the limited biological samples used for silver-stained 2DE analysis. Altogether, the protocol takes close to two days from first dimension separation to second dimension separation, followed by silver staining, scanning, and analysis.
authors.affiliation: Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, Serdang, Malaysia.
authors.forename: Yun Khoon
authors.initials: YK
authors.lastname: Liew
authors.forename: Vasanthakumari
authors.initials: V
authors.lastname: Neela
authors.forename: Rukman Awang
authors.initials: RA
authors.lastname: Hamat
authors.forename: Syafinaz Amin
authors.initials: SA
authors.lastname: Nordin
authors.forename: Pei Pei
authors.initials: PP
authors.lastname: Chong
mesh_terms: D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005399:Fishes; D006801:Humans; D057230:Limit of Detection; D008956:Models, Chemical; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D016622:Silver Staining / Q000379:methods*
chemical_list: D011506:Proteins