Stain efficiency and MALDI-TOF MS compatibility of seven visible staining procedures.
Visible stain is still the most popular protein staining method used in proteomic approaches. However, most published data have been derived from comparisons between visible dyes and fluorescent dyes. In this work, we have focused on seven widely used visible staining procedures--Neuhoff CCB, blue silver, and five silver stains (LKB SN, He SN, Yan SN, Vorum SN, and Blum SN)--and studied their stain efficiencies and MALDI-TOF MS compatibilities on 1-D and 2-D PAGE. It was concluded that blue silver is slightly better in terms of stain efficiency than Neuhoff CCB, but it presented worse MS compatibility. Neuhoff CCB presented better MS compatibility and superior linearity but worse sensitivity than silver stains. Among the five silvering procedures, He SN showed the best MS compatibility and a reasonable staining efficiency; Yan SN lowered the chances of obtaining the protein identity by PMF but gave the best stain efficiency; Vorum SN gave a very clear background and a great contrast, while Blum SN was the worst in this respect. The implications of these results for the selection of a convenient stain are discussed according to specific objectives as well as practical aspects.
Lin
Jian-feng
JF
Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen, 361005, China.
Chen
Qing-xi
QX
Tian
Hong-yu
HY
Gao
Xia
X
Yu
Mei-lan
ML
Xu
Gen-jun
GJ
Zhao
Fu-kun
FK
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003102:Colloids; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D011506:Proteins / Q000032:analysis*; D040901:Proteomics; D015203:Reproducibility of Results; D012394:Rosaniline Dyes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis*; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
D003102:Colloids; D011506:Proteins; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine; C004692:coomassie Brilliant Blue