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anti fbxo2 polyclonal rabbit antibody  (Proteintech)


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    Proteintech anti fbxo2 polyclonal rabbit antibody
    Anti Fbxo2 Polyclonal Rabbit Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/fbxo2+rabbit+polyclonal+antibody/FBXO2+Antibody/pm39487312-62-14-20
    Average 93 stars, based on 16 article reviews
    anti fbxo2 polyclonal rabbit antibody - by Bioz Stars, 2026-11
    93/100 stars

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    Related Articles

    Gene Expression:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    RNA Sequencing:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Expressing:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Activation Assay:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    In Vitro:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Marker:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Immunocytochemistry:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Fluorescence:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Organ Culture:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Cell Culture:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Immunoprecipitation:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Binding Assay:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Western Blot:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Negative Control:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Suspension:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Microarray:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Staining:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Labeling:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    In Situ Hybridization:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.

    Control:

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis
    Article Snippet: Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.Forty to 60 μg of tonsillar tissue lysates, CNA.42-immunoprecipitated proteins, or CEM/C2 (ATCC ® CRL-2264 ™) cell lysates were separated by SDS-polyacrylamide gels (90 V for 1 h) then transferred (100 V for 1 h) to nitrocellulose membranes.. The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).. After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.After washing, the membranes were incubated for 1 h at room temperature with HRP-goat anti-mouse IgM (1:2,000) or peroxidase conjugated goat anti-rabbit IgG (1:2,000) (Jackson Immunoresearch) and anti-GAPDH (1:10,000) to detect CNA.42, FBXO2, and GAPDH, respectively.



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    TaqMan gene expression assays used in the study (Thermo-Fisher Scientific Cat Number 4331182).

    Journal: Frontiers in Medicine

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis

    doi: 10.3389/fmed.2022.1013660

    Figure Lengend Snippet: TaqMan gene expression assays used in the study (Thermo-Fisher Scientific Cat Number 4331182).

    Article Snippet: The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).

    Techniques: Gene Expression

    RNA-Seq analysis of the PEAC cohort illustrates the differential correlation of the FDC genes associated with early perivascular and late mature developmental stages in the RA synovium. Strong positive correlations of the pericyte/fibroblast markers NG2, THY1 and αSMA with the PDGFR-β/PDGF-BB axis (A-I) , NG2 and FDC-CNA.42/FBXO2 (A-II) , and NG2, THY1 and αSMA (A-III) in RA. (B) Correlations of PDGF-BB and TNFα/LTβ with the expression of each other's receptors and early FDC developmental genes. PDGF-BB positively correlates with its receptor and the TNFα/LTβ receptors (B-I) , TNFα and LTβ negatively correlate with PDGFR-β expression and the early FDC markers NG2 and αSMA (B-II) . (C) Converse correlations of the PDGF-BB/PDGFR-β and the TNF-α/LT-β axes with the expression of mature FDC markers. PDGF-BB/PDGFR-β and TNF-α/LT-β differently correlate with the mature FDC related genes CXCL13 (B cell chemoattractant), BAFF (B cell survival factor), and antigen display and presentation to B cells namely complement receptors (CR1/CD35, CR2/CD21), and Fcg receptors (FcγRIIA/CD32A, FcγRIIB/CD32A). (D) Correlation of the RA synovial pathotypes with the expression of PDGF-BB, PDGFR-β, TNF-α, and LT-β. Person correlation coefficient (r) and adjusted p -values are shown with the corresponding plots and tables.

    Journal: Frontiers in Medicine

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis

    doi: 10.3389/fmed.2022.1013660

    Figure Lengend Snippet: RNA-Seq analysis of the PEAC cohort illustrates the differential correlation of the FDC genes associated with early perivascular and late mature developmental stages in the RA synovium. Strong positive correlations of the pericyte/fibroblast markers NG2, THY1 and αSMA with the PDGFR-β/PDGF-BB axis (A-I) , NG2 and FDC-CNA.42/FBXO2 (A-II) , and NG2, THY1 and αSMA (A-III) in RA. (B) Correlations of PDGF-BB and TNFα/LTβ with the expression of each other's receptors and early FDC developmental genes. PDGF-BB positively correlates with its receptor and the TNFα/LTβ receptors (B-I) , TNFα and LTβ negatively correlate with PDGFR-β expression and the early FDC markers NG2 and αSMA (B-II) . (C) Converse correlations of the PDGF-BB/PDGFR-β and the TNF-α/LT-β axes with the expression of mature FDC markers. PDGF-BB/PDGFR-β and TNF-α/LT-β differently correlate with the mature FDC related genes CXCL13 (B cell chemoattractant), BAFF (B cell survival factor), and antigen display and presentation to B cells namely complement receptors (CR1/CD35, CR2/CD21), and Fcg receptors (FcγRIIA/CD32A, FcγRIIB/CD32A). (D) Correlation of the RA synovial pathotypes with the expression of PDGF-BB, PDGFR-β, TNF-α, and LT-β. Person correlation coefficient (r) and adjusted p -values are shown with the corresponding plots and tables.

    Article Snippet: The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).

    Techniques: RNA Sequencing, Expressing

    Activation of sorted tonsillar stromal cell subsets with PDGF-BB and TNF-α/LT-β induces early and mature FDC markers in vitro . (A) The CD45 − tonsillar stromal subsets were sorted using combinations of NG2/αSMA, NG2/CNA.42, CNA.42/αSMA and CNA.42/CR2 Abs. (B) Type-1 Pericytes [NG2 + /αSMA + ; indicated by red * in (A,B) ], early FDCs; (CNA.42 + /NG2 + , CNA.42 + /αSMA + , CNA.42 + /CR2 − ; indicated in A and B by blue, magenta, and green *, respectively) and mature FDCs [CNA.42 + /CR2 + , indicated by brown * in (A,B) ] were treated with 300 ng/ml PDGF-BB or 100 ng/ml TNF-α+ 100 ng/ml LT-αβ and the fold change in FBXO2 (CNA.42), αSMA, Collagen 1, CR2, and FcγRIIB gene expression compared to untreated cells was calculated by Livak's DD equation and shown in bar graphs. (C) Treatment of the NG2 + /αSMA + type-1 pericyte subset with PDGF-BB in slide cultures for 6 days induced the expression of the FDC marker CNA.42 compared to untreated cells as demonstrated by Immunocytochemistry. Image J quantification of the mean fluorescence intensity (MFI) of CNA-42 of the different conditions is shown in the histogram. Data is representative of three different experiments and is expressed as the mean ± SEM.

    Journal: Frontiers in Medicine

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis

    doi: 10.3389/fmed.2022.1013660

    Figure Lengend Snippet: Activation of sorted tonsillar stromal cell subsets with PDGF-BB and TNF-α/LT-β induces early and mature FDC markers in vitro . (A) The CD45 − tonsillar stromal subsets were sorted using combinations of NG2/αSMA, NG2/CNA.42, CNA.42/αSMA and CNA.42/CR2 Abs. (B) Type-1 Pericytes [NG2 + /αSMA + ; indicated by red * in (A,B) ], early FDCs; (CNA.42 + /NG2 + , CNA.42 + /αSMA + , CNA.42 + /CR2 − ; indicated in A and B by blue, magenta, and green *, respectively) and mature FDCs [CNA.42 + /CR2 + , indicated by brown * in (A,B) ] were treated with 300 ng/ml PDGF-BB or 100 ng/ml TNF-α+ 100 ng/ml LT-αβ and the fold change in FBXO2 (CNA.42), αSMA, Collagen 1, CR2, and FcγRIIB gene expression compared to untreated cells was calculated by Livak's DD equation and shown in bar graphs. (C) Treatment of the NG2 + /αSMA + type-1 pericyte subset with PDGF-BB in slide cultures for 6 days induced the expression of the FDC marker CNA.42 compared to untreated cells as demonstrated by Immunocytochemistry. Image J quantification of the mean fluorescence intensity (MFI) of CNA-42 of the different conditions is shown in the histogram. Data is representative of three different experiments and is expressed as the mean ± SEM.

    Article Snippet: The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).

    Techniques: Activation Assay, In Vitro, Gene Expression, Expressing, Marker, Immunocytochemistry, Fluorescence

    Correlation of IL-6 expression with synovial pathotypes and FDC markers. (A) Boxplots displaying the correlation of synovial IL-6, JAK2, STAT1, and blood IL-6 expression with synovial pathotypes. (B,C) Correlation blots of synovial IL-6, IL-6R, JAK1, and JAK2 expression with the FDC markers complement receptor 1 (CR1/CD35) and CXCL13, respectively. Correlation of STAT3 and STAT1 with CR1 and CXCL13 respectively are also shown. (D) Correlation of synovial IL-6 expression with the markers associated with early FDC differentiation including NG2 (pericytes), αSMA (myofibroblasts), and FBXO2 (CNA.42). (E) IL-6 release from synovial organ and fibroblast cultures stimulated with 300 ng/ml PDGF-BB for 24 hrs and 6 days respectively. (E-I) Synovial organ culture showing a piece of synovial tissue placed in cell culture inserts mounted in 24-well plates (Upper). Diagrammatic representation of the synovial organ culture setup (S = Synovial Tissue, M = Culture Medium, F = Filter Device). (E-II) Rheumatoid arthritis synovial fibroblasts (RASFs) at base line (Day 1 = D1) and after 6-day (D6) stimulation with PDGF-BB. IL-6 levels at baseline and after stimulation are shown in (E-III) . Cultures were run in triplicates and data is expressed as the mean ± SEM. Significance was calculated using 2-tailed unpaired student T test and the p -value between baseline and PDGF-BB stimulation is shown.

    Journal: Frontiers in Medicine

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis

    doi: 10.3389/fmed.2022.1013660

    Figure Lengend Snippet: Correlation of IL-6 expression with synovial pathotypes and FDC markers. (A) Boxplots displaying the correlation of synovial IL-6, JAK2, STAT1, and blood IL-6 expression with synovial pathotypes. (B,C) Correlation blots of synovial IL-6, IL-6R, JAK1, and JAK2 expression with the FDC markers complement receptor 1 (CR1/CD35) and CXCL13, respectively. Correlation of STAT3 and STAT1 with CR1 and CXCL13 respectively are also shown. (D) Correlation of synovial IL-6 expression with the markers associated with early FDC differentiation including NG2 (pericytes), αSMA (myofibroblasts), and FBXO2 (CNA.42). (E) IL-6 release from synovial organ and fibroblast cultures stimulated with 300 ng/ml PDGF-BB for 24 hrs and 6 days respectively. (E-I) Synovial organ culture showing a piece of synovial tissue placed in cell culture inserts mounted in 24-well plates (Upper). Diagrammatic representation of the synovial organ culture setup (S = Synovial Tissue, M = Culture Medium, F = Filter Device). (E-II) Rheumatoid arthritis synovial fibroblasts (RASFs) at base line (Day 1 = D1) and after 6-day (D6) stimulation with PDGF-BB. IL-6 levels at baseline and after stimulation are shown in (E-III) . Cultures were run in triplicates and data is expressed as the mean ± SEM. Significance was calculated using 2-tailed unpaired student T test and the p -value between baseline and PDGF-BB stimulation is shown.

    Article Snippet: The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).

    Techniques: Expressing, Organ Culture, Cell Culture

    The FDC mAb CNA.42 recognizes FBXO2. (A) Immunoprecipitation (IP) and characterization of the CNA-42 binding protein. (A-I) Western blotting of total cell lysate (a), negative control (agarose beads only) and the CNA.42-immunoprecipitated proteins (c and d, 1.5 and 6 uls/lane respectively) from tonsillar single cell suspension probed with CNA-42. A single band is detectable at 120 Kd. (A-II) the reactivity of the CNA.42 mAb on the HuProt™ human proteome microarray showing subarray 9-1 of array 1300017931 (used for the CNA.42) with fluorescence detection at 633 nm excitation (a) and 543 nm excitation (b). (a) Staining with biotinylated anti-GST and Streptavidin-647. Rows 1-28 show generic staining of the GST-tagged immobilized human proteins, among them FBXO2 in row 11. (b) Probing with CNA.42 and Cy3 labeled anti-mouse IgM shows one hit, the human protein FBXO2 in the subarray. (A-III) Western blotting of tonsillar lysates with FBXO2 and CNA-42-specific antibodies recognize 120 Kd bands in the lysates [CNA.42 BP = CNA.42 binding protein]. (B) In situ hybridization of FBXO2 mRNA (green) showing intracellular signal in tonsillar CD21 + FDC reticula (red). (C) Western blotting of lysates from the CAN.42 expressing CEM cell line using mAb CAN.42 and anti FBXO2. CEM were untreated or treated either with Accell human FBXO2 siRNA (1 uM), or non-targeting control (NTC). GAPDH is used as a loading control. Compared to untreated cells, densitometric analysis with Image J indicates that FBXO2 siRNA-treated cells expressed 50% (*) and 35% (**) less FBXO2 and CNA.42, respectively.

    Journal: Frontiers in Medicine

    Article Title: Follicular dendritic cell differentiation is associated with distinct synovial pathotype signatures in rheumatoid arthritis

    doi: 10.3389/fmed.2022.1013660

    Figure Lengend Snippet: The FDC mAb CNA.42 recognizes FBXO2. (A) Immunoprecipitation (IP) and characterization of the CNA-42 binding protein. (A-I) Western blotting of total cell lysate (a), negative control (agarose beads only) and the CNA.42-immunoprecipitated proteins (c and d, 1.5 and 6 uls/lane respectively) from tonsillar single cell suspension probed with CNA-42. A single band is detectable at 120 Kd. (A-II) the reactivity of the CNA.42 mAb on the HuProt™ human proteome microarray showing subarray 9-1 of array 1300017931 (used for the CNA.42) with fluorescence detection at 633 nm excitation (a) and 543 nm excitation (b). (a) Staining with biotinylated anti-GST and Streptavidin-647. Rows 1-28 show generic staining of the GST-tagged immobilized human proteins, among them FBXO2 in row 11. (b) Probing with CNA.42 and Cy3 labeled anti-mouse IgM shows one hit, the human protein FBXO2 in the subarray. (A-III) Western blotting of tonsillar lysates with FBXO2 and CNA-42-specific antibodies recognize 120 Kd bands in the lysates [CNA.42 BP = CNA.42 binding protein]. (B) In situ hybridization of FBXO2 mRNA (green) showing intracellular signal in tonsillar CD21 + FDC reticula (red). (C) Western blotting of lysates from the CAN.42 expressing CEM cell line using mAb CAN.42 and anti FBXO2. CEM were untreated or treated either with Accell human FBXO2 siRNA (1 uM), or non-targeting control (NTC). GAPDH is used as a loading control. Compared to untreated cells, densitometric analysis with Image J indicates that FBXO2 siRNA-treated cells expressed 50% (*) and 35% (**) less FBXO2 and CNA.42, respectively.

    Article Snippet: The membranes were probed overnight at 4°C with 2 μg/mL of CNA.42 or 1:200 FBXO2 rabbit polyclonal antibody (Proteintech).

    Techniques: Immunoprecipitation, Binding Assay, Western Blot, Negative Control, Suspension, Microarray, Fluorescence, Staining, Labeling, In Situ Hybridization, Expressing, Control