Approximately 25% of patients who undergo kidney transplantation develop HLA-specific antibodies, the strength of which has not been previously correlated with graft failure. The strength of these de novo antibodies is investigated in this study. Serial dilution of strong HLA-specific allo-antibodies (up to 1:25,600) and testing with HLA-antigen-coated beads showed that the titer of the reaction to different HLA antigens is directly correlated to maximum fluorescence values and the molecules of equivalent soluble fluorochrome (MESF) values obtained by Luminex machines. Thus, the strength of antibodies can be measured utilizing maximum fluorescence and MESF. The strength of antibodies in the sera from 39 patients who subsequently had graft failure were markedly higher than those in the sera of 26 patients who continued to have good graft function (p = 0.0084). A clear increase in the strength of antibodies was identified in nine patients with a subsequent increase in serum creatinine levels. If analyzed for donor specificity, a strong association was noted for donor-specific MESF and failure (p = 0.00000027). Our results suggest that it is important to monitor the strength of antibodies when evaluating patient sera posttransplant.