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3 protocols using anti il17a antibody

1

Quantifying IL-17A in Psoriatic Skin

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Psoriatic and normal tissues were lysed in a protein lysis buffer (Intron, Daejeon, Korea). The concentration of protein was measured by a BCA Protein Assay Reagent (Pierce Biotechnology, Rockford, IL, USA). Samples were separated using sodium dodecyl sulfate-polyacrylamide gels, then transferred to nitrocellulose membranes. After blocking with skim milk, membrane was incubated with anti-IL-17A antibody (Abcam, Cambridge, MA, USA) or anti-actin antibody (Sigma, St. Louis, MO, USA). Membrane was then incubated with peroxidase-conjugated secondary antibodies. Western bands were visualized using enhanced chemiluminescence (Intron).
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2

Anti-Inflammatory Effects of XFBD

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XFBD was provided by Tianjin Modern TCM Innovation Center (TRT 200302). LPS was purchased from Sigma-Aldrich (Shanghai, China). Dexamethasone was purchased from Yuanye Bio (Shanghai, China). Anti-TNF-α antibody, anti-F4/80 antibody, anti-Neutrophil Elastase antibody, anti-IL17A antibody and Alexa Fluor®488-conjugated goat anti-Rat IgG were purchased from Abcam (Cambridge, MA, USA). Recombinant human IL17A was purchased from R&D systems (Minneapolis, USA). IL-17A monoclonal antibody (IL17A mAb) and mouse IgG1 kappa isotype control (IgG1K) were purchased from eBioscience (San Diego, California, USA). Anti-PD-1 antibody was purchased from Proteintech (Wuhan, China). HRP-conjugated goat anti-rabbit IgG was purchased from ZSGB-BIO (Beijing, China). Enzyme linked immunosorbent assay kits of TNF-α was purchased from ZCi BiO (Shanghai, China). glycyrrhizic acid, DMSO, RIPA lysis, BCA Protein Assay Reagent Kit and PMSF were purchased from Solarbio (Beijing, China). DAB substrate kit was purchased from Boster Biological (Wuhan, China). The myeloperoxidase (MPO) detection kit was purchased from Nanjing Jiancheng Bioengineering Institute.
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3

Histological and Immunohistochemical Analysis of Cardiac Tissue

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The excised hearts were fixed with 4% formalin for histological and immunohistochemical examinations. Following 24–48 h of dehydration, clearing, and embedding in paraffin wax, sliced Sects. (4 mm) were stained with haematoxylin and eosin and Masson’s trichrome and cardiac sections were dewaxed, subjected to antigen retrieval, and incubated with anti-IL17A antibody and anti-FOXp3 antibody (Abcam, Cambridge, UK), followed by incubation with a secondary antibody, and then analyzed using Image J (NIH V1.8.0.112).
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