Goat anti rabbit or donkey anti mouse igg hrp conjugated secondary antibodies

Cells were lysed in RIPA buffer with EDTA (Boston BioProducts) supplemented with protease (Roche) and phosphatase inhibitors (Pierce). Cleared lysate concentrations were obtained by DC Protein Assay (BioRad). Lysates were subjected to SDS-PAGE and transferred to nitrocellulose membranes (GE Healthcare). Western blots were conducted using: rabbit anti-Abl (Cell Signaling Technologies [CST], #2862); rabbit anti-pCrkL (CST #3181); mouse anti-CrkL (CST #3182); rabbit anti-EGFR (Abcam, Clone EP38Y); and mouse anti-β-actin (Sigma, Clone AC-74). Goat anti-rabbit or donkey anti-mouse IgG HRP-conjugated secondary antibodies (GE Healthcare) were used with chemiluminescence substrates (Pierce). Densitometry was assessed by ImageJ (5 (link),20 (link)).

Cells were lysed in boiling buffer with EDTA (Boston BioProducts) supplemented with 1X protease and 1% phosphatase inhibitor prepared following the manufacturer protocols (Sigma-aldrich, Cat.No. 11697498001 and P5726). Cleared lysate concentrations were obtained by a DC Protein Assay (BioRad). Lysates 30-40 μg were run on SDS-PAGE gels and transferred to nitrocellulose membranes (GE Healthcare). Western blots were conducted using: the Abcam antibodies to EGFR (Cat. No. 52892) and perforin (Cat. No. ab180773), and Cell Signaling Technology (CST) antibodies to GAPDH (Cat. No. 5174), JAK1 (Cat. No. 3332), STAT1 (Cat. No. 14994), p-STAT1 Y701 (Cat. No. 9167), PCAF/KA2B (Cat. No. 3378), Granzyme B (Cat. No. 4275), NFKB p65 (Cat. No. 82420), p-NFKB P65 (Cat. No. 3031S), and HSPB1 (Cat. No. 2402S). Goat anti-rabbit or donkey anti-mouse IgG HRP-conjugated secondary antibodies (GE Healthcare) were used with chemiluminescence substrates (Pierce). Densitometry was measured using ImageJ (v1.48).