Human/Mouse/Rat XIAP Antibody

Detection of Human and Mouse XIAP by Simple Western^TM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and C2C12 mouse myoblast cell line, loaded at 0.2 mg/mL. A specific band was detected for XIAP at approximately 59 kDa (as indicated) using 5 µg/mL of Goat Anti-Human/Mouse/Rat XIAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8221) 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.

Western Blot Shows Human XIAP Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and XIAP knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat XIAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8221) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for XIAP at approximately 52 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Rat XIAP by Immunohistochemistry

Oxycodone differentially induces pro-and anti-apoptotic signaling in cell bodies and axons. a Immunohistochemical analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF in striatum of rats treated with water (W) or oxycodone (O). Arrows point to axonal fascicles. Scale bar denotes 20 μm for all images. b Graphs of the densitometric analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF staining in bundles of the striatum rats treated with water (open bars) or oxycodone (grey filled bars). The graphs represent relative density measured as the ratio of the mean values of intensities obtained from oxycodone exposed tissue to the bundle intensities in striatum of water-exposed rat brains set as 1 (± SEM). Data obtained from three animals for each treatment. Statistical analysis was performed using Student’s t-test. eIF2 alpha -P, p < 0.001; ATF4, p < 0.001; Bax, p < 0.001; cleaved caspase 3, p < 0.001; XIAP, p < 0.001; and NF, p = 0.3. c Graphs of the densitometric analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF staining in cell bodies in striatum of rats treated with water (open bars) or oxycodone (grey filled bars). Data presented as a percent of positively stained cells relative to total number of cells. Data obtained from three animals for each treatment, Data expressed as mean value (± SEM). Statistical analysis of positively stained cells in oxycodone relative to water exposed striatum was performed using Student’s t-test. Phospho-eIF2 alpha, p < 0.001; ATF4, p = 0.2; Bax, p < 0.05; and XIAP, p < 0.01. d Western blot analysis of XIAP and phospho(Thr181)-Tau in rat cortex lysates. Left panel, the representative images of western blots of XIAP, phospho-Tau, total Tau, actin and GAPDH in cortex of rats treated with water (W) or oxycodone (O). Right panel, graphs of the densitometric analysis of XIAP and phospho-Tau western blots. The graphs represent the mean ratio of signal of XIAP to actin and phospho-Tau to total Tau. Oxycodone data were normalized to water samples in corresponding tissues set as 1 (± SEM). XIAP, n = 3, p < 0.05; P-Tau, n = 4, p < 0.003. Open bars—XIAP; and grey filled bars—P-Tau Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29571287), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Rat XIAP by Western Blot

Oxycodone differentially induces pro-and anti-apoptotic signaling in cell bodies and axons. a Immunohistochemical analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF in striatum of rats treated with water (W) or oxycodone (O). Arrows point to axonal fascicles. Scale bar denotes 20 μm for all images. b Graphs of the densitometric analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF staining in bundles of the striatum rats treated with water (open bars) or oxycodone (grey filled bars). The graphs represent relative density measured as the ratio of the mean values of intensities obtained from oxycodone exposed tissue to the bundle intensities in striatum of water-exposed rat brains set as 1 (± SEM). Data obtained from three animals for each treatment. Statistical analysis was performed using Student’s t-test. eIF2 alpha -P, p < 0.001; ATF4, p < 0.001; Bax, p < 0.001; cleaved caspase 3, p < 0.001; XIAP, p < 0.001; and NF, p = 0.3. c Graphs of the densitometric analysis of phospho-eIF2 alpha, ATF4, Bax, cleaved caspase 3 (Casp3), XIAP, and NF staining in cell bodies in striatum of rats treated with water (open bars) or oxycodone (grey filled bars). Data presented as a percent of positively stained cells relative to total number of cells. Data obtained from three animals for each treatment, Data expressed as mean value (± SEM). Statistical analysis of positively stained cells in oxycodone relative to water exposed striatum was performed using Student’s t-test. Phospho-eIF2 alpha, p < 0.001; ATF4, p = 0.2; Bax, p < 0.05; and XIAP, p < 0.01. d Western blot analysis of XIAP and phospho(Thr181)-Tau in rat cortex lysates. Left panel, the representative images of western blots of XIAP, phospho-Tau, total Tau, actin and GAPDH in cortex of rats treated with water (W) or oxycodone (O). Right panel, graphs of the densitometric analysis of XIAP and phospho-Tau western blots. The graphs represent the mean ratio of signal of XIAP to actin and phospho-Tau to total Tau. Oxycodone data were normalized to water samples in corresponding tissues set as 1 (± SEM). XIAP, n = 3, p < 0.05; P-Tau, n = 4, p < 0.003. Open bars—XIAP; and grey filled bars—P-Tau Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29571287), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of XIAP by Western Blot

Evaluation of apoptosis regulating proteins after lexatumumab (1 µg/ml) and DTIC (50 µg/ml) treatment in FEMX-1 and HHMS.The cells were harvested 24 h after treatment exposure and analyzed for various apoptotic markers. (A) Immuno-blot detection of Bcl-w, Bcl-XL, Mcl-1, Bak, Bax, Bik, Bad, BimEL, BimL and BimS expression in FEMX-1 cells. (B) Immuno-blot detection of cIAP-1, cIAP-2, XIAP, survivin and livin in FEMX-1 and HHMS cells. beta -actin is used as loading control. Cell lysates were prepared from three independent biological experiments, and representative blots are included in the figure. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23029050), licensed under a CC-BY license. Not internally tested by R&D Systems.