human mouse ciap pan specific antibody  (R&D Systems)

Journal: Frontiers in immunology

Article Title: Lipopolysaccharide sensitizes the therapeutic response of breast cancer to IAP antagonist.

doi: 10.3389/fimmu.2022.906357

Figure Lengend Snippet: FIGURE 5 MyD88 associates with and stabilizes cIAP proteins. MDA-MB-231 cells at sub-confluence were treated with LPS (100 ng/ml) or its combination with SM-164 (3 nM) −/+ TLR4i (5 mM) (A) and were treated with LPS (100 ng/ml) −/+ MyD88 inhibitor 4210 (10 mM) (B) for 8 h; 20 mg of whole- cell lysate protein from each sample was used to test protein levels of MyD88, cIAP1/2, and b-actin by Western blotting (WB). The quantified WB data were from five (A) or three (B) repeats. *p < 0.05 and **p < 0.01 between the indicated groups. (C) Cell lysate protein measuring 300 mg from samples, as in panel B, was immunoprecipitated with anti-MyD88 antibody −/+ 10 mM of MyD88 inhibitor 4210. In addition, one sample was incubated with normal IgG. The pulldown proteins were used to test cIAP1/2 and MyD88 by WB. The experiments were repeated three times with similar results.

Article Snippet: Purchase information of reagents are listed as follows: APExBIO (Boston, MA, USA), SM-164 (#A8815), AT-406 (#A3019), and BV6 (#B4653); EMD Millipore (Burlington, MA, USA), TLR4 inhibitor TAK-242 (#243984-11-4); eBioscience (San Diego, CA, USA), Annexin V (AnnV) Apoptosis Detection kit including AnnV antibody and propidium iodide (#88-8005-72); R&D Systems (Minneapolis, MN, USA), TNFa (R&D Systems Cat# 210- GMP) and pan cIAP antibody (#MAB3400); Proteintech (Rosemont, IL, USA), MyD88 antibody (#23230-1-AP); SigmaAldrich (St. Louis, MO, USA), b-actin antibody (#A5441), Protein Affinity gel (#P6486), Protein G affinity gel (#E3403), and LPS (L2630); and Invitrogen (Carlsbad, CA, USA), human TNF-a ELISA kit (#KHC3011).

Techniques: Western Blot, Immunoprecipitation, Incubation