Quantifying Anti-dsDNA Antibodies in Serum

Serum samples collected at necropsy were analyzed for anti‐dsDNA antibodies by ELISA as described earlier [49 (link), 53 (link)]. Briefly, pGEM‐3zf plasmid DNA (Promega) was digested with S1‐nuclease (Thermo Fisher Scientific) and biotinylated using Photoprobe Biotin (Vector Labs) following the manufacturer's instructions. Biotinylated dsDNA (1 ng/mL) was used for the assay.

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Venkatadri R., Sabapathy V., Dogan M., Mohammad S., Harvey S.E., Simpson S.R., Grayson J.M., Yan N., Perrino F.W, & Sharma R. (2022). Targeting Bcl6 in the TREX1 D18N murine model ameliorates autoimmunity by modulating T‐follicular helper cells and germinal center B cells. European Journal of Immunology, 52(5), 825-834.

Publication 2022

pGEM‐3zf plasmid DNA S1‐nuclease Photoprobe Biotin

Anti dsdna antibodies Assay Biotin Dsdna Elisa Necropsy Pgem Plasmid Promega Serum Vector

Corresponding Organization : University of Virginia

Other organizations : Wake Forest University, The University of Texas Southwestern Medical Center

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Variable analysis

independent variables

Serum samples collected at necropsy

dependent variables

Anti-dsDNA antibodies measured by ELISA

control variables

Biotinylated dsDNA (1 ng/mL) used for the assay

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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