title: Advanced negative detection method comparable to silver stain for SDS-PAGE separated proteins detection.
abstract: In order to achieve an easy, rapid and sensitive protocol to detect proteins in polyacrylamide gel, an advanced negative detection method comparable to silver stain is described. When a gel was incubated with Phloxine B and followed by the development in acidic solution, the zones where forming protein-dye complex were selectively transparent, unlike opaque gel background. Within 50 min after electrophoresis, down to 0.1-0.4 ng of gel-separated proteins (similar with silver stain) could be observed, without labor-intensive and time-consuming procedure. Comparing with the most common negative stain method, Imidazole-zinc stain, Phloxine B stain has been shown higher sensitivity and distinct contrast between the transparent protein bands/spots and opaque background than those; furthermore, it is no longer necessary to concern about retention time of observation. This technique may provide a sensitive and practical choice for proteomics researches.
authors.affiliation: School of Pharmacy, Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Pharmacy & Research Institute of Drug Development, Chonnam National University, Gwangju, South Korea.
authors.forename: Sun-Young
authors.initials: SY
authors.lastname: Hwang
authors.affiliation: School of Pharmacy, Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Wei-Tao
authors.initials: WT
authors.lastname: Cong
authors.affiliation: School of Pharmacy, Wenzhou Medical College, Wenzhou, Zhejiang, China.
authors.forename: Li-Tai
authors.initials: LT
authors.lastname: Jin
authors.affiliation: College of Pharmacy & Research Institute of Drug Development, Chonnam National University, Gwangju, South Korea. Electronic address: jkchoi@chonnam.ac.kr.
authors.forename: Jung-Kap
authors.initials: JK
authors.lastname: Choi
mesh_terms: D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D010697:Eosine I Bluish / Q000737:chemistry*; D006801:Humans; D016622:Silver Staining / Q000379:methods
chemical_list: D010697:Eosine I Bluish
keywords: Background stain; Negative stain; Phloxine B; Protein detection; SDS-PAGE