Human/Mouse BID Antibody

Detection of Human BID by Western Blot. Western blot shows Jurkat human acute T cell leukemia cell line treated with apoptosis inducer anti-Fas for the indicated times. PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for BID at approximately 20 kDa (as indicated). For additional reference short (5 seconds, left panel) and long (60 seconds, right panel) exposures are shown. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Detection of Human and Mouse BID by Western Blot. Western blot shows lysates of SVEC4-10 mouse vascular endothelial cell line and Jurkat human acute T cell leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for BID at approximately 20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human BID by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for BID at approximately 29 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse BID Antigen Affinity-purified Polyclonal Antibody (Catalog # AF860) 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 BID by Western Blot Survivin is predominantly expressed in obese hASCs and is influenced by the inflammatory environment. (a) Survivin protein expression and quantification in total adipose tissue (AT), adipocyte fraction (AD) and stromal-vascular fraction (SVF) from SAT of lean and obese patients. Fumarylacetoacetase (FAA) was used as a loading control. Ponceau S staining is also provided. *P<0.001 versus total AT and AD. (b) Survivin and p53 protein levels and quantification in hASCs from lean and obese patients. GAPDH was used as a loading control. *P<0.01 versus lean. (c) Survivin secreted levels and quantification in medium from lean- and obese-derived hASCs. Ponceau S staining was used to check loading. *P<0.05 versus lean conditioned medium. (d) Total cell extracts of lean- and obese-derived hASCs were subjected to immunoblotting with antibodies against apoptotic and antiapoptotic markers and GAPDH was used as a loading control. No significant differences were found. (e) Lean hASCs were cultured in RPMI medium (C: control) or for 24 h in conditioned medium (CM) from macrophages (M0, M1 and M2) and cells were subjected to immunobloting with survivin and GAPDH antibodies. *P<0.05 versus CM from control, and M0; #P<0.05 versus M2. (f) Left panel, lean hASCs were cultured in RPMI medium (C: control) or RPMI supplemented with IL-1 beta (IL1B) or IL-1 beta and IL-1 beta receptor antagonist (a); right panel, with CM of M1 macrophages or CM of M1 and IL-1 beta receptor antagonist (a). Survivin protein levels were measured and GAPDH was used as a loading control. Quantification of three experiments is shown. *P<0.05 versus control (c); #P<0.05 as indicated in figure. Data information: Values are expressed as mean±S.E.M. Statistical analysis: unpaired Student’s t-test. n=3–4 patients for each group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28518147), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human BID by Western Blot Survivin is predominantly expressed in obese hASCs and is influenced by the inflammatory environment. (a) Survivin protein expression and quantification in total adipose tissue (AT), adipocyte fraction (AD) and stromal-vascular fraction (SVF) from SAT of lean and obese patients. Fumarylacetoacetase (FAA) was used as a loading control. Ponceau S staining is also provided. *P<0.001 versus total AT and AD. (b) Survivin and p53 protein levels and quantification in hASCs from lean and obese patients. GAPDH was used as a loading control. *P<0.01 versus lean. (c) Survivin secreted levels and quantification in medium from lean- and obese-derived hASCs. Ponceau S staining was used to check loading. *P<0.05 versus lean conditioned medium. (d) Total cell extracts of lean- and obese-derived hASCs were subjected to immunoblotting with antibodies against apoptotic and antiapoptotic markers and GAPDH was used as a loading control. No significant differences were found. (e) Lean hASCs were cultured in RPMI medium (C: control) or for 24 h in conditioned medium (CM) from macrophages (M0, M1 and M2) and cells were subjected to immunobloting with survivin and GAPDH antibodies. *P<0.05 versus CM from control, and M0; #P<0.05 versus M2. (f) Left panel, lean hASCs were cultured in RPMI medium (C: control) or RPMI supplemented with IL-1 beta (IL1B) or IL-1 beta and IL-1 beta receptor antagonist (a); right panel, with CM of M1 macrophages or CM of M1 and IL-1 beta receptor antagonist (a). Survivin protein levels were measured and GAPDH was used as a loading control. Quantification of three experiments is shown. *P<0.05 versus control (c); #P<0.05 as indicated in figure. Data information: Values are expressed as mean±S.E.M. Statistical analysis: unpaired Student’s t-test. n=3–4 patients for each group Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28518147), licensed under a CC-BY license. Not internally tested by R&D Systems.