cy5 rabbit anti vimentin  (Bioss)

Journal: Respiratory Research

Article Title: Increased susceptibility to organic dust exposure-induced inflammatory lung disease with enhanced rheumatoid arthritis-associated autoantigen expression in HLA-DR4 transgenic mice

doi: 10.1186/s12931-022-02085-8

Figure Lengend Snippet: Expression of lung autoantigens, serum inflammatory markers, and autoantibodies following ODE exposure. A Confocal images of lung tissue from treatment groups stained for citrulline (CIT)- and malondialdehyde-acetaldehyde (MAA)-modified proteins as well as vimentin, with nuclei staining by DAPI (blue). Images were analyzed using Zen 2012 software (Zeiss). Bar graphs depict mean integrated density (+ SD) of B CIT- or C MAA-modified proteins, and D vimentin, quantified per each mouse. E R 2 -values demonstrating co-localization of MAA-CIT, CIT-Vimentin, and MAA-Vimentin within each treatment group. Panels F – H demonstrate serum levels of the murine acute phase reactant protein, pentraxin-2 ( F ), serum levels of IgG antibody to MAA-modified proteins utilizing human serum albumin, collagen type II, and vimentin as the substrate antigen ( G ), and BALF levels of IgG antibody to MAA ( H ), all quantified by ELISA. Statistical differences versus saline-treated mice are indicated by # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001. Brackets between groups represents significant gene-ODE interaction ( P-int) and “a” (p < 0.05) is WT-ODE vs. DR4-ODE. RU: relative units. Line scale is 100 μm

Article Snippet: Paraffin embedded lung sections from mice were rehydrated and subjected to antigen retrieval using citrate buffer and steam, and then blocked with goat serum and incubated with either Cy5 rabbit anti-vimentin (Bioss, Woburn, MA, 1:100), Zenon AF 594 label (Invitrogen, Carlsbad, CA), rabbit polyclonal IgG antibody to MAA, or a mouse monoclonal anti-peptidyl-citrulline antibody (clone F95 IgMκ, Millipore Sigma, Burlington, MA).

Techniques: Expressing, Staining, Modification, Software, Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: Increased susceptibility to organic dust exposure-induced inflammatory lung disease with enhanced rheumatoid arthritis-associated autoantigen expression in HLA-DR4 transgenic mice

doi: 10.1186/s12931-022-02085-8

Figure Lengend Snippet: ODE-induced lung autoantigens, vimentin, serum inflammatory markers, and autoantibodies in the setting of CIA in DR4 mice. A Confocal images of lung tissue from treatment groups stained for citrulline (CIT)- and malondialdehyde-acetaldehyde (MAA)-modified proteins as well as vimentin. Bar graphs in B – D depict integrated density (+ SD) of B CIT- or C MAA-modified proteins, and D vimentin, quantified per each mouse. Bar graphs in E – G demonstrate serum levels of murine pentraxin-2 ( E ), serum levels of IgG antibody to MAA-modified proteins ( F ) utilizing human serum albumin, collagen type II, and vimentin as the substrate antigen, and BALF levels of IgG antibody to MAA ( G ), all quantified by ELISA. Statistical differences versus saline-treated control mice are designated by # p < 0.05, ## p < 0.01, #### p < 0.0001. Lines denote differences between groups (*p < 0.05, **p < 0.01, ****p < 0.0001). RU: relative units. Line scale is 100 μm

Article Snippet: Paraffin embedded lung sections from mice were rehydrated and subjected to antigen retrieval using citrate buffer and steam, and then blocked with goat serum and incubated with either Cy5 rabbit anti-vimentin (Bioss, Woburn, MA, 1:100), Zenon AF 594 label (Invitrogen, Carlsbad, CA), rabbit polyclonal IgG antibody to MAA, or a mouse monoclonal anti-peptidyl-citrulline antibody (clone F95 IgMκ, Millipore Sigma, Burlington, MA).

Techniques: Staining, Modification, Enzyme-linked Immunosorbent Assay