Microwave-enhanced ink staining for fast and sensitive protein quantification in proteomic studies.
A novel microwave-enhanced ink staining method was developed for rapid and sensitive estimation of protein content in sample buffers containing chaotropes, dyes, detergents, and reducing agents. Dye-based Blue-Black ink was used to quantitatively visualize proteins spotted on a nitrocellulose membrane. The total staining time was greatly reduced to 3 min by brief exposure to microwave radiation. The stained membrane was washed with distilled water, baked in a microwave oven for complete desiccation, transparentized with mineral oil, and documented by a desktop scanner or densitometer. Only 1 microL of protein sample (protein solubilized in SDS-PAGE sample buffer or IEF rehydration buffer) was used for protein spotting. The novel solid-phase protein assay gives a 500-fold dynamic range from 19.5 to 10000 ng/microL and can be scaled up for high-throughput protein quantification analysis. The fast, sensitive and low-cost microwave-enhanced ink staining procedure is ideal for protein quantification in proteomic analysis.
Wu
Xue-Ping
XP
Laboratory of Molecular Biology and MOE Laboratory of Protein Science, Department of Biological Science and Biotechnology, Tsinghua University, Beijing 100084, China.
Cheng
Yong-Sheng
YS
Liu
Jin-Yuan
JY
D000818:Animals; D002417:Cattle; D003101:Collodion / Q000737:chemistry; D003720:Densitometry; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007091:Image Processing, Computer-Assisted; D007281:Ink; D008872:Microwaves; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry; D013997:Time Factors
D003902:Detergents; D012709:Serum Albumin; D003101:Collodion