Rabbit anti ftl

Western blot was performed as described previously (22 (link)). The primary reagents were used: rabbit anti-FTL (dilution, 1:1,000; Abcam, UK), rabbit anti-GAPDH antibodies (dilution, 1:1,000; Cell Signaling Technology, Inc., USA), horseradish peroxidase (HRP)–conjugated anti-rabbit IgG (dilution, 1:2,000; Cell Signaling Technology, Inc., USA), ECL Chemiluminescent HRP Substrate (Merck Millipore, USA), and BCA Protein Assay Kit (Beijing Biosynthesis Biotechnology Co., China). The gray levels of bands were analyzed by ImageJ software. The relative gray level was represented by relative IntDen (relative IntDen = the integrated band density (IntDen) of FTL/IntDen of GAPDH).

Standard Western blots were undertaken using RIPA (radioimmunoprecipitation assay) lysates as previously described [16 (link)]. A total of 10–20 ug of lysate was resolved via SDS-PAGE. Primary antibodies were as follows: anti-vinculin (1:5000) (Cat#4650, Cell Signalling Technology, Danvers, MA, USA), mouse anti-β-actin (1:5000) (Cat#sc-69879, Santa Cruz Biotechnology, Dallas, TX, USA), rabbit anti-TGM2 (1:1000) (Cat#ab2386, Abcam, Cambridge, UK), rabbit anti-FTL (Cat#ab69090, Abcam, Cambridge, UK), rabbit anti-ANXA6 (Cat#31026, Abcam, Cambridge, UK), mouse anti-MVP (Cat#ab97311, Abcam, Cambridge, UK), rabbit anti-NDRG1 (Cat#5196S, Cell Signalling Technology, Danvers, MA, USA), and rabbit anti-SOD2 (Cat#13194S, Cell Signalling Technology, Danvers, MA, USA). HRP-conjugated goat anti-mouse (Cat#1706516, Bio-Rad, Hercules, CA, USA) and goat anti-rabbit (Cat#1706515, Bio-Rad, Hercules, CA, USA) secondary antibodies were used at 1:5000 and 1:3000, respectively.