Sequencing & Spectrometry Tools | Protein Research
Mass Spectrometry Analysis Tools:
FOCUS™ FASTsilver™ produces crystal clear backgrounds and maximal peptide recovery needed for critical analysis by mass spectrometry.
For mass spectrometry analysis, complete proteolytic digestion and recovery of peptides is required for optimal analysis, however silver ions in traditional silver staining kits inhibit proteolytic digestion. In addition, glutaraldehyde, a common sensitizer in silver stains, modifies peptide lysine residues preventing complete digestion and recovery.
FOCUS™ FASTsilver™ produces high quality silver staining without the use of glutaraldehyde and is supplied with a highly efficient silver ion removal reagent, SilverOUT™. SilverOUT™ removes silver ions, which permits complete peptide digestion and extraction of peptides for maximal recovery.
A reliable method for the proteolytic digestion of proteins in gel for subsequent analysis by mass spectrometry.
The protein spots are first excised from the gel and transferred to a proteomic grade tube (see accessories). Silver stained gel pieces are washed with SilverOUT™ to remove inhibitory silver ions and then the gel pieces are treated with our proprietary Trypsin-Digestion Buffer Mix, a mixture of MSG-Trypsin™ and an optimal digestion buffer, which ensures reliable and efficient protein digestion.
The resulting digested peptides are extracted with Pep-Extract™, a high diffusion peptide extraction buffer. The extracted peptides are suitable for mass spectrometry analysis without any subsequent treatments or cleaning procedures.
A 96-well format kit suitable for processing larger numbers of protein spots concurrently and compatible with spot-picking instruments. Fewer than 96 samples a batch may also be processed without any waste.
Excise protein spots and transfer to the supplied proteomic grade titer plate, destain silver stained gel pieces with SilverOUT™ and then incubate with our proprietary Trypsin-Digestion Buffer Mix, a mixture of MSG-Trypsin™ and an optimal digestion buffer, which ensures reliable and efficient protein digestion.
The resulting digested peptides are extracted with Pep-Extract™, a high diffusion peptide extraction buffer. The extracted peptides are suitable for mass spectrometry analysis without the need for further cleaning procedures.
Trypsin is a serine endopeptidase that specifically cleaves peptide bonds on the carboxy side of s-aminoethyl cysteine, arginine and lysine residues. Typically there is little or no cleavage at arginyl-proline and lysyl-proline bonds.
Trypsin undergoes autolysis, producing trypsin fragments that interfere with sequence analysis. G-Biosciences' MSG-Trypsin™ is a chemically modified trypsin that is enzymatically active and yet resistant to autolysis. MSG-Trypsin™ is methylated, TPCK treated and quality tested for mass spectrometry.
Unlike other trypsin preparations, MSG-Trypsin™ is highly stable, maintaining its activity in severe denaturing buffers and as a result, is shipped without requiring dry ice and can be stored for a long period without any loss of activity.