Anti rabbit igg hrp

Anti-CD47-APC, anti-CD47-FITC, anti-CD11c-APC-Vio770, anti-CD11-PE, anti-CD14-FITC, anti-CD80-PE, anti CD83-PE, anti-CD86-PE, anti-HLA-DR-FITC, anti-HLA-ABC-FITC, anti-Hsp70-FITC, and 7-Amino-Actinomycin D (7-AAD) fluorescent DNA dye were purchased from Miltenyi. All monoclonal antibodies (mAbs) used in flow cytometry experiments were used at 1:200 titer unless otherwise specified. Anti-calreticulin-FITC mAb (EPR3924; 1:50) and anti-GAPDH were from Abcam. Anti-mouse/human/rat CD47 mAb or mouse IgG isotype control were purchased from Bio X Cell. Anti-phospho-EGFR (Y1608), anti-EGFR, and anti-phospho-Akt (S473) were from Cell Signaling Technology. Secondary antibodies anti-rabbit IgG-HRP or anti-mouse IgG+IgM+IgA-HRP were from Bethyl Laboratories. Gefitinib was purchased from Selleckchem, and its IC₅₀ was determined for each cell line using Cell-Counting Kt-8 (Dojindo Laboratories) according to the manufacturer's instructions. Gefitinib IC₅₀ for each cell line, reported in Table 1, was used for all the experiments. Recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) was purchased from Miltenyi, and interferon (IFN)-α (IntronA) from SP Europe.

The following antibodies were used: the anti-p-NFκB p65 (Ser536) (clone 9H31) and the anti-NFκB p65 (clone C22B4) from Cell Signaling Technology (Danvers, MA, USA); the anti-Gapdh (clone 6C5) from Santa Cruz Biotechnology (Dallas, Texas, USA); the anti-embryonic Myosin Heavy Chain, eMyHC (clone F1.652) from Developmental Studies Hybridoma Bank (Iowa City, IA, USA); polyclonal anti-Laminin from Sigma-Aldrich; the anti-CD45 PE/Cy5 conjugated (clone 30-F11) from BD Pharmigen™ (San Jose, CA, USA); the anti-CD31 Alexa Fluor 488 conjugated (clone MEC13.3) from Biolegend (San Diego, CA, USA); the anti-mouse IgG TRITC conjugated and the anti-rabbit IgG TRITC conjugated from Sigma Aldrich; the anti-mouse IgG Alexa Fluor 488 conjugated from Invitrogen; the anti-rabbit IgG Chromeo 488 conjugated from Abcam (Cambridge, MA, USA); the anti-mouse IgG and the anti-rabbit IgG HRP conjugated from Bethyl Laboratories (Montgomery, TX, USA).

Antibodies used in this study are as follows: anti-Vps34 (Cell signaling, #4263), anti-Beclin1 (Cell signaling, #3738; Santa Cruz, sc-48341 for immunoprecipitation), anti-ATG14L (MBL, PD016), anti-UVRAG (Cell signaling, #13115), LC3B (Cell signaling, #2775), α-tubulin (Santa Cruz, sc-23948), anti-vinculin (SIGMA, V9131) anti-Flag HRP conjugate (Sigma, A8592), anti-HA HRP conjugate (Cell signaling, #2999S) and the secondary antibodies conjugated with HRP, anti-Rabbit IgG-HRP (Bethyl, #A120-101P) and anti-Mouse IgG-HRP (Bethyl, #A90-116P). The affinity resins for the protein purification or immunoprecipitation were obtained from SIGMA (Flag-M2 agarose, A2426), GE Healthcare (GSH-Sepharose, 17-0756-01; Protein G-Sepharose, 17-0618-01), and Invitrogen (Protein A-Sepharose, #101041). Chloroquine (C6628), α-pinene (147524), Δ-3-carene (115576), α-cedrol (93483), β-caryophyllene (22075), and α-humulene (5375) were obtained from Sigma-Aldrich. High glucose (25 mM) DMEM (LM-001-07) and glucose-free DMEM (LM001-56) were from Welgene.