Human cIAP-2/HIAP-1 Antibody

cIAP-2/HIAP-1 in Human Colon Tissue. cIAP-2/HIAP-1 was detected in immersion fixed paraffin-embedded sections of human colon tissue using Goat Anti-Human cIAP-2/HIAP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8171) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to colon glands. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human cIAP‑2/HIAP‑1 by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human cIAP-2/HIAP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8171) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for cIAP-2/HIAP-1 at approximately 68 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.

Detection of Human cIAP‑2/HIAP‑1 by Simple Western^TM. Simple Western lane view shows lysates of Raji human Burkitt's lymphoma cell line and Daudi human Burkitt's lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for cIAP-2/HIAP-1 at approximately 70 kDa (as indicated) using 5 µg/mL of Goat Anti-Human cIAP-2/HIAP-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8171) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human cIAP-2/HIAP-1 by Western Blot IAP expression in glioblastoma cell lines. Expression levels of cIAP1, cIAP2, XIAP and ML-IAP were analyzed by western blotting and quantified in U87MG and GL261 adherent GBM cell lines, and in GBM6 and GBM9 spheres. Expression level of beta -actin served as loading control. The four GBM cell lines expressed heterogeneously cIAP1, cIAP2, XIAP and ML-IAP. A representative experiment of four experiments is shown. Quantification was performed using ImageJ software (National Institutes of Health, Bethesda, MD, USA) and data presented were normalized to beta -actin expression Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27490930), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human cIAP-2/HIAP-1 by Immunocytochemistry/Immunofluorescence Prognostic value of cIAP1, cIAP2, XIAP and ML-IAP protein expression in human glioblastomas (cohorts 1 and 2). (a) cIAP1-, cIAP2-, XIAP- and ML-IAP-positive stainings in GBM. IAPs were heterogeneously expressed by tumor cells in GBM samples (Table 1). Stainings were diffused with a stronger punctuated positivity into the cytoplasm. Black arrows highlight cIAP2-positive nuclei. Scale bars, 50 μm. (b) Correlation of ML-IAP protein expression with PFS and OS in cohort 1. The cutoff was 35% and was determined by performing a ROC curve. ML-IAP expression of ⩾35% was correlated with a poor prognosis. (c) Correlation of ML-IAP protein expression with PFS and OS in cohort 2. The cutoff was the same as that for cohort 1 analysis (35%). ML-IAP expression of ⩾35% was correlated with a poor prognosis Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27490930), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human cIAP-2/HIAP-1 by Western Blot Apoptosis and IAP expression upon SMAC mimetic GDC-0152 treatment in glioblastoma cell lines. (a) Apoptosis (SubG0/G1) of DMSO control and GDC-0152-treated cells was determined by flow cytometry of propidium iodide-stained nuclei and percentage of apoptosis is shown. U87MG and GL261 cell lines were treated for 72 h and GBM6 and GBM9 cell lines were treated for 8 days at the indicated concentrations. At these respective time points, percentage of U87MG cells dead by apoptosis, percentage of GL261 cells, percentage of GBM6 cells and percentage of GBM9 cells. Data are expressed as mean+S.E.M. Three independent experiments were performed for the GL261 cell lines and five for the U87MG, GBM6 and GBM9 cell lines. *P<0.05; **P<0.01; ***P<0.005. (b) Expression levels of cIAP1, cIAP2, XIAP and ML-IAP were analyzed by western blotting. Cell lines were treated with 1 μM of GDC-0152. U87MG and GL261 were treated for 72 h and GBM6 and GBM9 cell lines for 8 days. In all GBM cell lines GDC-0152 decreased IAP expression. Expression level of beta -actin served as loading control. A representative experiment of three experiments is shown Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27490930), licensed under a CC-BY license. Not internally tested by R&D Systems.