Mouse anti-Human pTau231 Monoclonal Antibody (Clone KT231)

Cross reactivity test: Microtiter wells were coated with various peptides and recombinant proteins. KT231 was used as the primary antibody and peroxidase conjugated goat anti-mouse IgM was used as the secondary antibody. Result: KT231 does not cross-react with Tau peptide or recombinant proteins without any phosphorylation site.

ELISA: Microtiter wells were coated with KT231 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 (KT231) was used as the detection antibody. Result: KT231 and KT232 can be used as a matched antibody pair to detect and quantify the concentration of human pTau231 with high specificity.

Western Blot: Synthetic pTau peptide fragments containing phosphorylated threonine 231 were crosslinked with BSA (phospho T231). The conjugates were run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. Peptide with phosphorylated threonine 217 or threonine 181 or without any phosphorylation site were processed with the same procedure as negative controls. KT231 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgM was used as the secondary antibody. Tau (phospho T231) 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: KT231 can specifically detect recombinant human Tau (phospho T231) by Western blotting.