A dilution series of two TAMRA labeled peptides were bound to a PVDF membrane using a dot blot apparatus. Peptide #1 is conjugated directly to TAMRA via an amide bond whereas peptide #2 contains an aminohexanoyl linker between the TAMRA and the peptide. Also note that the peptides differ in sequence and in length, peptide #1 has 15 amino acids and peptide #2 has 29 amino acids. A dilution series for each peptide was made starting with approximately 0.5 nmoles of peptide and followed by a series of 1:2 dilutions. After binding of the peptides to the PVDF membrane, the membrane was then blocked in 5% milk in 1X TBST for 1 hour at room temperature. The membrane was then incubated in with the ab9093 antibody at a 1:2000 dilution in 5% milk in 1X TBST for 1 hour at room temperature. Following incubation with the primary antibody the blot was then washed 3 times in 1X TBST and then incubated with a 1:10,000 dilution of an anti-mouse-HRP secondary antibody for 45 minutes at room temperature