Human ADAM10 Ectodomain Antibody

Detection of ADAM10 Ectodomain by Western Blot

Short chain fatty acids (SCFA) mildly increase amyloidogenic processing.(A) Western blot analysis and (B) its densitometry quantification of 3 months old brain homogenates of control (Ctrl)- and SCFA-supplemented germ-free (GF) APPPS1 animals. The A beta level is significantly increased in SCFA group in comparison to Ctrl, despite unaffected APP FL levels. APP CTFs show a decreased C83 (**) to C99 (*) ratio. We could not detect alterations in protein levels of secretases involved in APP processing (ADAM10, BACE1, and gamma-secretase/PSEN1). m = ADAM10 mature form; im = ADAM10 immature form. Data represent mean ± SEM (unpaired T-test; n(Ctrl) = 3, n(SCFA) = 3). (C) Upper panel: gamma-Secretase reconstituted into lipid vesicles was incubated at 37°C together with the C99-based substrate C100-His6 in the presence of increasing doses of a SCFA mixture (375 and 750 μM final concentration of total SCFA of an equimolar mixture of Na-acetate, Na-propionate, and Na-butyrate) for 24 hr. Production of A beta was analyzed by immunoblotting. gamma-Secretase inhibitor (GSI) L-685,458 (0.4 μM) was used as a negative control. No alterations in A beta levels were detected in the presence of SCFA. Lower panel: Qualitative analysis of individual A beta species via Tris‐Bicine‐Urea SDS‐PAGE reveals that SCFA treatment does not alter the ratio among the different A beta species (A beta37-38-40-42/43) suggesting no direct effects on modulation of gamma-secretase cleavage. (D) Aggregation kinetics of monomeric A beta40 recorded by the increase in fluorescence of Thioflavin T incubated with either 30 mM NaCl (Ctrl) or 30 mM SCFA mixture do not show any significant difference, suggesting that SCFA do not directly modify A beta fibrillarization. Data points represent mean ± SD from three independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33845942), licensed under a CC-BY license. Not internally tested by R&D Systems.