HPLC Method for Analysis of IRDye® 800CW NHS Ester on BIST A Column

Separation type: Bridge Ion Separation Technology, or BIST™ by SIELC Technologies

IRDye® 800CW NHS Ester is an infrared dye used for labeling primary and secondary amine residues in proteins. Using SIELC’s newly introduced BIST™ method, this dye can be retained on a negatively-charged, cation-exchange BIST™ A+ column. There are two keys to this retention method: 1) a multi-charged, positive buffer, such as TMEDA formate, which acts as a bridge, linking the negatively charged dye to the negatively-charged column surface and 2) a mobile phase consisting mostly of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Tartrazine can be separated, retained, and detected at 600 nm.