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Our Abpromise guarantee covers the use of ab92773 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab92773 was created by mutagenesis of the P9 serine residue (Ser338) on the reactive center loop to cysteine. This then provided a free thiol group for incorporation of N,N'-dimethyl-N-(acetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) (NBD), a fluorescent probe highly sensitive to changes in solvation and hydrophobic environment. The fluorescence emission of P9-NBD PAI1 is enhanced 6-7 fold upon insertion of the reactive center loop into beta-sheet A following complex formation with proteinases, formation of the latent species, or cleavage by elastase. The incorporated probe is excited at 480 nm and displays a broad emission spectrum with a peak centered 542 nm with a resultant blue-shift to 520 nm following reactive center loop insertion. The modified PAI1 is nearly as active as wt PAI-1 and is more resistant to the spontaneous latency reaction making this an excellent tool for monitoring reaction rates of PAI1. ab92773 has been utilized in a number of studies to determine the rates of loop insertion and SERPIN reaction mechanisms when reacted with various proteinases, inactivating antibodies and conformational changes imposed by the binding of vitronectin.
Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.
Constituents: 0.1M Sodium chloride, 0.05M Sodium phosphate, 1mM EDTA, pH 6.6
This product is an active protein and may elicit a biological response in vivo, handle with caution.
ab92773 has not yet been referenced specifically in any publications.