HEp-2 Immunofluorescence Assay Protocol

HEp-2 immunofluorescence assays were performed as previously described [22 (link)]. In brief, serum was diluted 1/200 and then applied to HEp-2 slides (Antibodies Incorporated or BD). Staining was detected using goat anti-mouse IgG FITC at 1:500 (Southern Biotech) ANAs were scored on intensity of nuclear and cytoplasmic staining on a scale of 0–3 and for the presence or absence of mitotic chromatin using wide-field fluorescence microscopy (Olympus IX83; Cellsense software). The scorer used blinded to the genotype and gender of the mice.

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Gordon R.A., Tilstra J.S., Marinov A., Nickerson K.M., Bastacky S.I, & Shlomchik M.J. (2020). Murine lupus is neutrophil elastase-independent in the MRL.Faslpr model. PLoS ONE, 15(4), e0226396.

Publication 2020

goat anti-mouse IgG FITC

Anas Anti igg Antibodies Chromatin Cytoplasmic Fitc Fluorescence microscopy Genotype Goat Immunofluorescence assays Mice Serum

Corresponding Organization : University of Pittsburgh

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Variable analysis

independent variables

Genotype of mice
Gender of mice

dependent variables

Intensity of nuclear and cytoplasmic staining on a scale of 0-3
Presence or absence of mitotic chromatin

control variables

Serum dilution (1/200)
Staining detection (goat anti-mouse IgG FITC at 1:500)
Wide-field fluorescence microscopy (Olympus IX83; Cellsense software)
Scorer blinded to genotype and gender of mice

Annotations

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