Anti human igg4

Human recombinant HGF was purchased from GenScript (Nanjing, China). The c-MET kinase inhibitors JNJ38877605 and rapamycin (RAPA) were obtained from Selleckchem (Houston, TX). Antibodies used include c-MET mAb, p-MET (Tyr^1234/1235) mAb, Notch mAb, HES-1 mAb, and GAPDH mAb. Antibodies used in this study include c-MET mAb, p-MET (Tyr1234/1235) mAb (Abcam, U.S.A.), Notch mAb (Abcam, U.S.A.), HES-1 mAb (Abcam, U.S.A.), GAPDH mAb (Abcam, U.S.A.), Anti-mouse IgG (Abcam, U.S.A.), anti-CTLA4, #EPR1476 (Abcam, U.S.A.) and anti-human IgG4(Abcam, U.S.A.). All antibodies were purchased from Cell Signaling Technology, Inc.

Antibodies used for flow cytometry analysis were: anti-human/mouse CD45 (Invitrogen, # 17-0451-82), anti-human CD20 (Invitrogen, #25-0209-42), anti-human MARCH3 (Invitrogen, #PA5-60351), biotin-anti-B2M (Abcam, #ab269365), anti-human DTWD1 (Invitrogen, #MA5-27492), anti-human IgG1 (Abcam, #ab99776), anti-human RPS12 (Abcam, #ab175219) and anti-human IgG4 (Abcam, #ab1930).
The detailed staining procedure of flow cytometry analysis has been described previously (13 (link)). Briefly, nonspecific binding was blocked with Fcγ receptor blocker (Invitrogen, #14-9161-73). For surface staining, cells were incubated on ice with fluorescent-coupled antibodies for 15 min and then washed twice with cold PBS. For intracellular staining, cells were treated using a Cytofix/Cytoperm kit (BD, #554717) according to manufacturer’s introductions and then incubated at 4°C with the relevant fluorescent-coupled antibodies for 15 min. After two washes with cold PBS, cells were re-suspended with PBS/0.5%BSA/2mM EDTA. Samples were then measured with a flow cytometer (Thermal Fisher Attune NxT).