Figure 3: ELISA assay of the interaction of PiPE, mycerial wall component elicitor (MWC) and mycerial homogenate (MH) from P. infestans with Histagged protein from the full length of calcium-dependent protein kinase (CDPK) from potato cv. Rishiri (R1-gene) (RiCDPK2), consisting of kinase-I to X with junction- and Ca2+-binding-domain (KJC). The wells were coated with 250 ng of fusion KJC. The respective wells were treated with 250 ng of fusion 38 kD elicitor, MWC, FH, and detected with 50 ul of mouse monoclonal anti-PiPE antibodies-2A11 (PiPE-Abs) diluted to 1:2000 in phosphate buffered saline with 1% BSA and 0.05% NaN3. The readings were taken at A405 nm after 1 hr of substrate addition. KJC was added with PiPE, and the His-tagged protein from the full length of RiCDPK2 interacted with His-PiPE; KJC was added with HWC, and the His-tagged protein from KJC interacted with HWC; and KJC was added with FH, and fusion protein from KJC interacted with the mycelial homogenate. PiPE, MWC, and MH detected with PiPE-Abs served as a positive control. Experiments were repeated two times. Similar results were obtained. Each bar is the mean ± SE of two replicates.