Mouse anti-Human pTau217 Monoclonal Antibody (Clone KT225)

Cross reactivity test: Microtiter wells were coated with various peptides and recombinant proteins. KT225 was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Result: KT225 does not cross-react with Tau peptide or recombinant proteins without any phosphorylation site.

ELISA: Microtiter wells were coated with KT225 at 3 ug/mL as the capture antibody. Synthetic peptides with different phosphorylation sites were used as the antigen. Peroxidase conjugated mouse anti-human Tau monoclonal antibody (KT233) was used as the detection antibody. Result: KT225 and KT233 can be used as matched antibody pair to detect and quantify the concentration of human pTau217 with high specificity.

Western Blot: Synthetic pTau peptide fragments containing phosphorylated threonine 217 were crosslinked with BSA (phospho T217). The conjugates were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. Peptide with phosphorylated threonine 231 or threonine 181 or without any phosphorylation site were processed with the same procedure as negative controls. KT225 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Tau (phospho T217) band was visualized using ECL Western Blotting Substrate. Lane 1: 25 ng of Tau (phospho T217)-BSA conjugate Lane 2: 25 ng of Tau (phospho T231)-BSA conjugate Lane 3: 25 ng of Tau (phospho T181)-BSA conjugate Lane 4: 25 ng of Tau Result: KT225 can specifically detect Tau (phospho T217) by Western blotting.