Tnt quick transcription translation system

The following antibodies were used for immunoblotting: NT-1 recognizing the PS1-NTF [61] (link), α-loop (Chemicon) raised against PS1-CTF, N1660 (Sigma) raised against the C-terminal of nicastrin, 3891 (ProSci Inc.) raised against Pen-2 and α-GAPDH (Acris GmbH) recognizing GAPDH. For the quantification of secreted Aβ38, 40 and 42 with Meso Scale Discovery (MSD) technology, C-terminal specific antibodies (Aβ 1-x) were used and detection was performed by SULFO-TAG™ 6E10 antibody. For quantification of sectreted Aβ40 and Aβ43 using Aβ40 Wako II ELISA kit (Wako Chemicals GmbH) and FL 1-43 ELISA kit (Immuno-biological Laboratories), respectively, the capture antibody was BNT77 for Aβ40 and Aβ38-43 for Aβ43. Detection antibodies were BA27 (Aβ40) and 82E1 (Aβ43), respectively. Unless otherwise stated, all chemicals were from Sigma–Aldrich. Plasmid pGEM1, TNT® Quick transcription/translation system, and deoxynucleotides were purchased from Promega and ³⁵S-Met from PerkinElmer. All enzymes were obtained from Fermentas except Phusion DNA polymerase that was from Finnzymes. Oligonucleotides were from Eurofins MWG Operon.