Human/Mouse cIAP Pan-specific Antibody

Detection of Human/Mouse cIAP by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line transfected with human cIAP-1 or human cIAP-2. PVDF membrane was probed with 0.5 µg/mL Mouse Anti-Human/Mouse cIAP Pan-specific Monoclonal Antibody (Catalog # MAB3400) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, lysates of Raji human Burkitt's lymphoma cell line, Jurkat human acute T cell leukemia cell line, and SW3T3 mouse contact inhibited fibroblast cell line were included. Specific bands for cIAP-1 and cIAP-2 were detected at approximately 80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human cIAP by Simple Western^TM. Simple Western lane view shows lysates of HEK293 human embryonic kidney cell line transfected with either cIAP-1 or cIAP-2 and Raji human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for cIAP-1 and cIAP-2 at approximately 73 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse cIAP Pan-specific Monoclonal Antibody (Catalog # MAB3400). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human cIAP (pan) by Western Blot cIAPs and c-FLIP exert an efficient double brake against TLR3-mediated apoptosis in normal bronchial epithelial cells.a Primary human bronchial epithelial cells (HBEC) were exposed to Poly(I:C) (100 μg/ml) for 24 h, and the secretion of chemokines/cytokines were determined. Error bars represent S.E.M. of two independent experiments. b Primary HBEC were exposed to Poly(I:C) (100 μg/ml) for 6 and 24 h (left panel), or to cisplatin for 24 h (right panel), and the percentage of Annexin V+ cells was determined by flow cytometry. Error bars represent S.E.M. of three (left panel) and two (right panel) independent experiments. c Human immortalized bronchial epithelial HBEC3-KT cells were treated with Poly(I:C) (100 μg/ml) for 6 h and 24 h (left panel), or to cisplatin (100 μM) for 24 h (right panel), and the percentage of Annexin V+ cells was determined. Error bars represent S.E.M. of four (left panel) and three (right panel) independent experiments. **P < 0.01. d HBEC3-KT cells were transfected with a control non-silencing siRNA (siNS) or targeting TLR3 (siTLR3), exposed to Poly(I:C) (100 μg/ml) for 24 h, and the secretion of chemokines/cytokines was determined by ELISA. Error bars represent S.E.M. of three independent experiments. *P < 0.05. e HBEC3-KT cells were transfected as in d, lysed, and immunoblotted as indicated. f The indicated cells were lysed and immunoblotted for TLR3 and actin. Representative images of three independent experiments. g HBEC3-KT cells transfected with a control siRNA (siNS) or targeting c-FLIP (siFLIP), and were treated for 1 h with BV6 (5 μM). The cells were then exposed for 6 h to increasing doses of Poly(I:C), and the percentage of Annexin V+ cells was determined. Error bars represent S.E.M. of four independent experiments. *P < 0.05 and **P < 0.01. h HBEC3-KT cells were treated as in f, lysed, and immunoblotted as indicated. i HBEC3-KT transfected with siNS or siFLIP were then exposed to Poly(I:C) (100 μg/ml) for the indicated times in presence or absence of BV6 (5 μM). The cells were then lysed and immunoblotted as indicated. Caspase cleavage products are indicated by arrowheads. Representative images of two independent experiments Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30158588), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human cIAP (pan) by Western Blot cIAPs and c-FLIP exert an efficient double brake against TLR3-mediated apoptosis in normal bronchial epithelial cells.a Primary human bronchial epithelial cells (HBEC) were exposed to Poly(I:C) (100 μg/ml) for 24 h, and the secretion of chemokines/cytokines were determined. Error bars represent S.E.M. of two independent experiments. b Primary HBEC were exposed to Poly(I:C) (100 μg/ml) for 6 and 24 h (left panel), or to cisplatin for 24 h (right panel), and the percentage of Annexin V+ cells was determined by flow cytometry. Error bars represent S.E.M. of three (left panel) and two (right panel) independent experiments. c Human immortalized bronchial epithelial HBEC3-KT cells were treated with Poly(I:C) (100 μg/ml) for 6 h and 24 h (left panel), or to cisplatin (100 μM) for 24 h (right panel), and the percentage of Annexin V+ cells was determined. Error bars represent S.E.M. of four (left panel) and three (right panel) independent experiments. **P < 0.01. d HBEC3-KT cells were transfected with a control non-silencing siRNA (siNS) or targeting TLR3 (siTLR3), exposed to Poly(I:C) (100 μg/ml) for 24 h, and the secretion of chemokines/cytokines was determined by ELISA. Error bars represent S.E.M. of three independent experiments. *P < 0.05. e HBEC3-KT cells were transfected as in d, lysed, and immunoblotted as indicated. f The indicated cells were lysed and immunoblotted for TLR3 and actin. Representative images of three independent experiments. g HBEC3-KT cells transfected with a control siRNA (siNS) or targeting c-FLIP (siFLIP), and were treated for 1 h with BV6 (5 μM). The cells were then exposed for 6 h to increasing doses of Poly(I:C), and the percentage of Annexin V+ cells was determined. Error bars represent S.E.M. of four independent experiments. *P < 0.05 and **P < 0.01. h HBEC3-KT cells were treated as in f, lysed, and immunoblotted as indicated. i HBEC3-KT transfected with siNS or siFLIP were then exposed to Poly(I:C) (100 μg/ml) for the indicated times in presence or absence of BV6 (5 μM). The cells were then lysed and immunoblotted as indicated. Caspase cleavage products are indicated by arrowheads. Representative images of two independent experiments Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30158588), licensed under a CC-BY license. Not internally tested by R&D Systems.