A comprehensive evaluation of imidazole-zinc reverse stain for current proteomic researches.
In this paper, we comprehensively evaluated the capability of imidazole-zinc reverse stain (ZN) in comparative proteomics. Three commonly used protein gel staining methods, including silver (SN), SYPRO Ruby (SR), and CB stain were investigated alongside for comparison purpose. A transparency scanning procedure, which may deliver more even and contrasting gel images, was found best for documenting ZN stained gels. Our results showed that ZN was more sensitive than SN, SR, and CB. It may reveal as few as 1.8 ng of proteins in a gel. Moreover, ZN was found to provide a linear dynamic range of staining for revealing proteins up to 140 ng, and show an insignificant staining preference. To analyze a ZN stained 2-D gel image that generally comprises an apparent but even background, the Melanie 4 software was found more suitable than others. Furthermore, ZN demonstrated an equivalent or better MS compatibility than the other three staining methods. Intense and comprehensive MS profiles were frequently observed for ZN stained gel spots. Approximate two-third of ZN stained gel spots were successfully identified for protein identities. Taken together, our results suggest that the prompt, cost effective and versatile ZN is well suited for current proteomic researches.
Lin
Ching-Yu
CY
Graduate Institute of Applied Science and Engineering, Catholic Fu-Jen University, Taipei, Taiwan, Republic of China.
Wang
Vinchi
V
Shui
Hao-Ai
HA
Juang
Rong-Huay
RH
Hour
Ai-Ling
AL
Chen
Pei-Sing
PS
Huang
Hui-Ming
HM
Wu
Szu-Yu
SY
Lee
Jen-Chieh
JC
Tsai
Tzung-Lin
TL
Chen
Han-Min
HM
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007093:Imidazoles / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D015032:Zinc / Q000737:chemistry*
D007093:Imidazoles; C029899:imidazole; D015032:Zinc