A Gaussian curve for the binned frequencies of log2 ratios was determined using Igor Pro v6. 1 and the mean was subtracted from all log2 ratios to center the population of average AD research use control log2 ratio and control 1 control 2 at zero. The SD was determined via the Gaussian width as 0. 72 log2 ratio units for AD control and 0. 30 for the null experiment. A total of 144 proteins were considered significantly changed and met the follow ing criteria, i fell outside the null experiment distribution or beyond 99. 9% confidence interval, ii had an absolute value 1. 17, iii had a coefficient of variation of less than 100% and iv had a signal to noise ratio greater than 10 in both control measurements.

True bio marker FDR was estimated by counting the false positives surviving the above filters in the null experiment and cal culating this number as a percentage of the total number of biomarkers proposed in the list for the AD control comparison, as described in results. The list of gene sym bols for these proteins was Inhibitors,Modulators,Libraries input into DAVID pathway analysis v6. 7, and significantly changing ontological classes of proteins were further considered in the context of available references which related each class to AD. Antibodies Primary antibodies used in these studies were as follows, FITC conjugated CD41 integrin aIIb, Abcam, Cambridge, MA, USA, APC conjugated CD45, THBS1, beta actin, the antibody dilutions for THBS1 and beta actin reflect prior dilution Inhibitors,Modulators,Libraries of each antibody with glycerol. Immunoblotting Equal concentrations of protein from each sample were loaded into a 10% acrylamide gel and separated by SDS PAGE.

Proteins were transferred onto polyvinylidene fluoride Immobilon P membranes overnight at 4 C. Immunoblots were blocked for 2 hours at room temperature with Tris buffered saline Tween and blocking buffer and probed for the protein of interest with a primary antibody overnight at 4 C. The following day, blots were incubated with fluorophore Inhibitors,Modulators,Libraries conjugated Inhibitors,Modulators,Libraries sec ondary antibodies for 1 hour in the dark. All blots were scanned and quantified using the Odyssey Infrared Imaging System. Statistical analysis was performed using a two tailed Students t test. Results and discussion Participant selection Characteristics of participants with clinically diagnosed AD and controls are presented in Table 1. Controls were selected to be as similar as possible to AD patients.

As expected, there was a Inhibitors,Modulators,Libraries significant difference between the groups in MMSE scores. Aspirin status was matched between groups to help control for any effect of aspirin on the platelet proteome. By matching for aspirin usage, we were able to obtain a sample more representative of the general population affected by AD. Furthermore, ideal biomarkers will change in disease inde pendent of factors such MEK162 ARRY-162 as medications.