The Tech Manual says to use saliva from known secretors and non-secretors as controls. It also lists under "Reagents", "Human (polyclonal) anti-A and anti-B. Note: some monoclonal reagents may not be appropriate for use, therefore, apprpriate controls are essential." Saline is also run. The anti-A used is diluted till it produces a 2+ agglutination. For testing the A substance only, I would have 4 tubes: 1) known secretor's saliva +Anti-A, 2) known non-secretor's saliva + anti-A, 3) saliva being tested + anti-A, & 4) saline + anti-A. After these incubate, A1 cells are added to each tube to see if the A substance in the saliva neutralized the diluted anti-A. Now that I have thought it through, only the non-secretors would produce agglutination so in a way that is actually the positive control--at least in my brain.