Anti goat igg antibody sc 2020

Proteins (20 µg of cellular protein extract and 5 µL of concentrated culture medium) were separated on 10% SDS-PAGE, transferred on 0.22 µm nitrocellulose membranes (Trans-Blot® Turbo^TM Transfert Pack, Bio-rad) and then incubated with antibodies as previously described^{6 (link)}. The primary antibodies used for western blot analysis were: CLU (sc-6419), Bcl2 (sc-493), and GAPDH (sc-365062) antibodies from Santa Cruz Biotechnology; beclin-1 (#3738), LC3B (#2775), and cleaved caspase 3 (#9664) antibodies from Cell signaling; mono-and poly-ubiquitin antibody (BML-PW8810) from Enzo Life Sciences; P62 (610498) from BD Transduction Laboratories; β-actin antibody (A5316) from Sigma-Aldrich and sarcomeric-actin antibody (m0874) from Dako. The horseradish peroxidase-labeled secondary antibodies used were: anti-rabbit IgG (NA934V) and anti-mouse IgG (NA931) antibodies from GE healthcare and anti-goat IgG antibody (sc-2020) from Santa Cruz Biotechnology. The dilution of antibodies used for Western blot is provided for each sample analyzed (Supplementary Table 1). The Chemidoc® camera (Biorad) was used for imaging the membranes and densitometric measurements of the bands were analyzed with the Image Lab software (Bio-Rad).

Cells were lysed with the Triton X-100 lysis buffer (Sigma-Aldrich). The supernatants from whole-cell lysates were collected after centrifugation. Denatured proteins were subjected to 8% SDS-PAGE and blotted onto a polyvinylidene difluoride membrane. The membranes were blocked with 5% bovine serum albumin (BSA, Sigma-Aldrich), followed by probing with the following primary antibodies: Hsp90 (sc-69703), αM integrin (ICRF44) from IQ Products; β2 integrin (sc-8420), caspase-1 (sc-56036) and β-actin (sc-47778) from Santa Cruz; Hsp70 (ab2787) from Abcam. The membranes were then incubated with horseradish peroxidase-conjugated anti-mouse IgG antibody (sc-2055), or anti-goat IgG antibody (sc-2020) from Santa Cruz, and visualized with the ECL solution using the ChemiDoc MP system (Bio-Rad) according to the manufacturer’s protocol. β-actin was used as a positive control for sample normalization.