Dear Claire, Per our conversation this morning, please note that the timing for HMB45 Primary Antibody and Innovex Two-step Alk Phos was not enough instead of 10-10. Best results are obtained at 30 for antibody and 30 for second step Alk Phos. This information is in the data sheet. Zahra Naser, PH.D. Innovex Biosciences