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The reaction was then quenched by the addition of 5x reducing buffer (50% glycerol, 250 mM Tris-Cl [pH 6.9], 10% SDS, 0.05% bromophenol blue, 6.25% beta-mercaptoethanol), boiled and separated by sodium dodecyl sulfate polyacrylamide (SDS-PAGE) on 15% polyacrylamide gels

Amex will remain the preferred premium brand. The biggest risk to Visa/Mastercard is losing Cap One as a debit/credit card issuer or having to renegotiate terms, considering that Cap One is now in a very competitive position against those networks. Banks that see Cap One as a competitor will probably drop Pulse as their PIN network