Donkey anti rabbit igg hrp

Following antibodies were obtained from indicated distributors: anti-annexin A2 (rabbit monoclonal, (CST D11G2, Danvers, MA), anti-annexin A2 (mouse monoclonal 610068, BD biosciences), anti-caveolin-1 (rabbit polyclonal Sc-894, Santa Cruz), anti-cavin-1 (anti-PTRF, rabbit polyclonal AP7421a, Abgent, San Diego, CA), anti-S100A10 (mouse monoclonal 4E7E10, CST), anti-phosphotyrosine (mouse monoclonal 4G10, Merck Millipore, Darmstadt, Germany), anti-eNOS (mouse monoclonal 610296, BD Biosciences), anti-p-Src (Y418, BS4176) and anti-p-Src (Y529, BS4729) (rabbit polyclonal, Bioworld Technology, St. Louis Park, MN), Donkey anti-rabbit IgG HRP and anti-mouse IgG HRP (Promega). OLA (Wako), EPA (Nu-Chek Prep, Elysian, MN) and DHA (Cayman Chemicals, Ann Arbor, MI) were obtained from indicate distributors. Aprotinin (Wako), α-2-antiplasmin (Haematologic Technologies Essex Junction VT), 4-(2-Aminomethyl) benzensulfonyl fluoride (AEBSF, nacalai tesque, Kyoto Japan), chymostatin (Sigma-Aldrich, St. Louis, MO) and Gö6976 (LC Laboratories, Woburn, MA) were obtained from indicated distributors. Other reagents not specified were obtained from Wako.

TCA-precipitated supernatant fractions and pellets were analysed by SDS-PAGE and immunoblotting as described in Kebbi-Beghdadi et al. [36 (link)]. Proteins of interest were detected using a mouse monoclonal anti-V5 epitope antibody (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) (1/5000 dilution) or anti β lactamase antibody (VWR International GmbH, Dietikon, Switzerland) (1/1000 dilution) and MreB was detected using a home-made rabbit polyclonal antibody (1/5000 dilution). First antibodies were incubated overnight at 4 °C. Secondary antibodies, goat anti-mouse IgG-HRP (BioRad, Cressier, Switzerland) and donkey anti-rabbit IgG-HRP (Promega, Dübendorf, Switzerland), both diluted 1/3000 were applied during 1 h at room temperature. Immunoblots were revealed with ECL^TM Prime Western Blotting Detection Reagent (Amersham, GE Healthcare, Glattbrugg, Switzerland) and analysed on ImageQuant LAS4000 mini (Amersham, GE Healthcare, Glattbrugg, Switzerland).