Amersham ecl primer western blotting detection reagent

Left ventricular samples from the non-infarcted area or from the cells were collected, washed, and lysed with a RIPA solution (THERMOFISHER, USA) supplemented with PMSF (MERCK, USA). The proteins were then extracted, and the proteins concentrations were quantified by BCA^{52 (link)}. Proteins (35 μg) were denatured, separated by SDS-PAGE electrophoresis and transferred to a polyvinylidene difluoride (PVDF) membrane (MERCK MILLIPORE, USA). The transferred membranes were blocked using 5% bovine serum albumin (BSA) in TBST and incubated with the primary antibodies summarized in the supplementary material. Bands were visualized using enhanced chemiluminescent ECL (AMERSHAM ECL Primer Western Blotting Detection Reagent (GE HEALTHCARE, NJ, USA) (RPN2232)), using a ChemiDoc XRS + system with Image Lab software from BIO-RAD Laboratories (BERKELEY, CA, USA).

Further, 10 µg of protein from 4 h and 24 h schistosomula and 50 ng of MBP and MBP–SmSoxS1 protein were used for Western analysis. The custom SmSoxS1-1 primary antibody (GenScript, Piscataway, NJ, USA) was diluted to 0.68 µg/mL in 5% milk-PSBTw (PBS/0.1% Tween-20) solution and divided into primary antibody and peptide block solution. The peptide block solution contained the peptide for the SmSoxS1-1 antibody at a 10× concentration by weight. An HRP-link goat anti-rabbit secondary antibody (GE Healthcare, Chicago, IL, USA) was used at a 1:2000 dilution in 1% milk-PBSTw and visualized using Amersham ECL Primer Western Blotting Detection Reagent (GE Healthcare, Chicago, IL, USA). The membranes were exposed to autoradiography film and developed for 10 min.