Validation and in vivo characterization of research antibodies for Moesin, CD44, Midkine, and sFRP-1.

Background: A major goal of the Target Enablement to Accelerate Therapy Development for Alzheimer’s disease (TREAT-AD) program is develop and identify high-quality tools test target or mechanistic hypotheses. As part this initiative, it important that commercial reagents including research antibodies being used interrogate drug targets have confirmed validation data in knock-out cell lines. Ideally, these should also utility both in vitro vivo studies such levels proteins tissues can be quantified. Methods: We evaluated against TREAT-AD protein Moesin (Uniprot ID: P26038), CD44 P16070), Midkine P21741) Secreted frizzled-related 1, referred as “sFRP-1” (sFRP-1; Uniprot Q8N474). Moesin, sFRP-1, were selective based on Western blot analysis was compare brain homogenates from a mouse model with AD-relevant pathology (5XFAD) versus age-matched C57BL/6J control mice. Results: Anti-Moesin ab52490 reacted homogenate predicted molecular weight 68 kDa. expression 2.8 times higher 5xFAD compared WT. Anti-CD44 ab189524 band at size 82 1.9 Anti-Midkine AF7769 ~16 kDa 17.8 greater Anti-sFRP-1 ab267466 35 predicted. sFRP-1 11.9 WT. Conclusions: These confirm commercially available CD44, Midkine, mice provide insight into use 5XFAD engagement proteins.