Ultrasensitive fluorescence protein detection in isoelectric focusing gelsusing a ruthenium metal chelate stain

SYPRO Ruby IEF Protein Gel Stain is an ultrasensitive, luminescent stain op timized for the analysis of protein in isoelectric focusing gels. Proteins are stained in a ruthenium-containing metal complex overnight and then rins ed in distilled water for 2 h. Stained proteins can be excited by ultraviol et light of about 302 nm (UV-B transilluminator) or with visible light of a bout 470 nm, Fluorescence emission of the dye is maximal at approximately 6 10 nm. The sensitivity of the SYPRO Ruby IEF protein gel stain is superior to colloidal Coomassie blue stain and the highest sensitivity silver staini ng procedures available. The SYPRO Ruby IEF protein gel stain is suitable f or staining proteins in nondenaturing or denaturing carrier ampholyte isoel ectric focusing and immobilized pH gradient gel electrophoresis. The stain is compatible with N,N'-methylenebisacrylamide or piperazine diacylamide cr oss-linked polyacrylamide gels as well as with agarose gels and high tensil e strength Duracryl gels. The stain does not contain extraneous chemicals ( formaldehyde, glutaraldehyde, Tween-20) that frequently interfere with pept ide identification in mass spectrometry. Successful identification of stain ed proteins by peptide mass profiling is demonstrated.