Alexafluor 680 conjugated goat anti mouse igg

Dithiothreitol (DTT), ethylenediaminetetraacetic acid (EDTA), 45% glucose, kynurenic acid, proteinase K (PK), sodium chloride (NaCl), Tris-HCl and Triton X-100 were purchased from Sigma (St. Louis, MO). Millicell 6-well plate inserts (Biopore filter CM, PTFE inserts 30-mm diameter and 0.4-um pore size, Cat No. PICM03050), E. coli Rosetta DE3 competent cells, Overnight Express^TM Autoinduction System 1 (Cat. no. 71300-3) and BugBuster^® Master Mix (Cat No. 71456-4) were purchased from Millipore (Billerica, MA). Bradford protein assay kit was purchased from Bio-Rad Laboratories (Hercules, CA). Horse serum, penicillin, streptomycin, and low-melting agarose (Cat No. 15517-022) were purchased from Invitrogen (Carlsbad, CA). Anti-PrP mouse monoclonal antibodies POM1 and 6D11 were purchased from Prionatis AG (Schlieren, Switzerland) and Covance, respectively. Anti-β-actin antibody was purchased from Sigma-Aldrich. Alexa Fluor 680 conjugated goat anti-mouse IgG was purchased from Invitrogen. Goat anti-rabbit IR800 conjugated IgG was purchased from Rockland Immunochemicals (Gilbertsville, PA), propidium iodide (PI) from Molecular Probes (Cat No. P3566), and Halt™ Protease and Phosphatase Inhibitor Cocktail (Cat No. 78440) from Life Technologies. All other reagents were obtained from Thermo Fisher Scientific (Waltham, MA) unless otherwise mentioned.

Reagents were purchased from the following sources: Mouse antibody against S6 (#2317s) and rabbit antibodies against phospho S6-S240/244 (#5364) and Arg-II (#55003S) were purchased from Cell Signaling Technology; Mouse antibody against Tubulin (T5168) and Goat Serum Donor Herd (G6767) were from MilliporeSigma (Burlington, MA, United States); Mouse antibody against p16 (sc-81156) was purchased from Santa Cruz Technology Inc. (Dallas, United States); Secondary antibodies IRDye 800-conjugated affinity purified goat anti-rabbit IgG was purchased from BioConcept (Alschwil, Switzerland) and Alexa fluor 680-conjugated goat anti-mouse IgG was from Invitrogen (Lucerne, Switzerland); Mouse IgG blocking reagent (MKB-2213) was from Vector Laboratories (Peterborough, United Kingdom); Alexa Fluor 488 conjugated-Goat anti-mouse IgG(H + L) secondary antibody (A-11001) was from Thermo Fisher Scientific (Waltham, MA United States); Dihydroethidium (DHE) was from Molecular Probes/Invitrogen (Lucerne, Switzerland), and the membrane-permeable 4,5-diaminofluoresceine diacetate (DAF-2DA) was from VWR international SA (Dietikon, Switzerland).