Detecting dsDNA Autoantibodies in Mice

Detection of dsDNA autoantibodies in mouse sera was performed as described (19 (link)). In Short: 0.01% Poly-L-lysine (Sigma-Alderich) were used to precoat MaxiSorp plates (Nunc). After 2 h at RT, plates were coated with dsDNA from calf thymus (20µg/ml; Sigma-Aldrich) in H₂0 over night at 4°C. Sera were added in 1/3 serial dilutions starting at 1/20, pooled serum of SLE affected MRL/lpr mice, with a starting dilution of 1/250, served as standard. Goat-anti-mouse IgG (1 mg/ml) coupled to AP was used for detection.

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Szumilas N., Corneth O.B., Lehmann C.H., Schmitt H., Cunz S., Cullen J.G., Chu T., Marosan A., Mócsai A., Benes V., Zehn D., Dudziak D., Hendriks R.W, & Nitschke L. (2021). Siglec-H-Deficient Mice Show Enhanced Type I IFN Responses, but Do Not Develop Autoimmunity After Influenza or LCMV Infections. Frontiers in Immunology, 12, 698420.

Publication 2021

MaxiSorp plates dsDNA from calf thymus

Anti igg Autoantibodies Dilution Dsdna Goat Lysine Mouse Mrl lpr mice Poly Serum Thymus

Corresponding Organization : Friedrich-Alexander-Universität Erlangen-Nürnberg

Other organizations : Erasmus MC, Universitätsklinikum Erlangen, Technical University of Munich, Semmelweis University

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Variable analysis

independent variables

Concentration of Poly-L-lysine used for precoating plates
Concentration of dsDNA from calf thymus used for coating plates
Dilution of sera samples

dependent variables

Detection of dsDNA autoantibodies in mouse sera

control variables

Type of plates used (MaxiSorp plates from Nunc)
Incubation time and temperature for precoating and coating plates
Goat-anti-mouse IgG coupled to AP used for detection

positive control

Pooled serum of SLE affected MRL/lpr mice with a starting dilution of 1/250

negative control

Not explicitly mentioned

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