Journal: Arthritis Research & Therapy

Article Title: Effect of dsDNA binding to SmD-derived peptides on clinical accuracy in the diagnosis of systemic lupus erythematosus

doi: 10.1186/ar2266

Figure Lengend Snippet: Binding of SmD peptides to dsDNA. (a) SmD1, SmD3 and biotinylated control peptides (PM1-α and Ribosomal P) were serially diluted in dilution buffer (0.01–10 μg/ml) and incubated on dsDNA-coated ELISA plates. Unbound peptides were removed by washing. Immobilized peptides were detected by streptavidin–horseradish peroxidase conjugate in combination with 3,3',5,5'-tetramethylbenzidine substrate. The SmD1 peptide showed dsDNA binding starting with a concentration of approximately 40 ng/ml, and the SmD3 peptide starting with approximately 0.6 μg/ml. No dsDNA binding could be observed with PM1-α and Ribosomal P. (b) The inhibitory effect of the SmD1 and SmD3 peptides was analysed by testing an anti-dsDNA-positive sample from a systemic lupus erythematosus patient on dsDNA-coated microtitre strips preincubated with increasing concentrations of the SmD peptides as described above. Detection of bound human dsDNA antibodies was according to the instructions for use of the dsDNA ELISA (Dr Fooke Laboratories). No inhibition was observed for SmD3, but inhibition was observed for SmD1 starting at a concentration of approximately 1 μg/ml. OD, optical density.

Article Snippet: The Sm ELISA (catalogue number 25010; Dr Fooke Laboratorien GmbH) is based on native highly purified SmD antigen from a bovine source.

Techniques: Binding Assay, Control, Incubation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Inhibition

Journal: Arthritis Research & Therapy

Article Title: Effect of dsDNA binding to SmD-derived peptides on clinical accuracy in the diagnosis of systemic lupus erythematosus

doi: 10.1186/ar2266

Figure Lengend Snippet: Bridging experiment. A serum sample with high-titre anti-dsDNA antibodies but no anti-SmD1 (amino acids 89–119) reactivity was spiked with increasing concentrations of dsDNA (plasmid, also used in dsDNA ELISA; Dr Fooke Laboratories) and incubated for 30 minutes at room temperature. The dilution series was subsequently tested for anti-dsDNA and anti-SmD1 reactivity in ELISA according to the instructions for use of the respective kit. The anti-dsDNA reactivity as determined by ELISA significantly decreased with increasing concentrations of dsDNA. No decrease in anti-SmD1 reactivity could be observed (data not shown). Error bars indicate the x -fold standard deviation of the duplicate determinations.

Article Snippet: The Sm ELISA (catalogue number 25010; Dr Fooke Laboratorien GmbH) is based on native highly purified SmD antigen from a bovine source.

Techniques: Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Incubation, Standard Deviation

Journal: The Journal of Experimental Medicine

Article Title: Augmentation of Vα14 Nkt Cell–Mediated Cytotoxicity by Interleukin 4 in an Autocrine Mechanism Resulting in the Development of Concanavalin a–Induced Hepatitis

doi:

Figure Lengend Snippet: Production of IL-4 and IFN-γ from Con A–activated Vα14 NKT cells. Whole spleen cells from B6, Vα14 NKT KO, Vα14 NKT, and RAG KO mice (A) or the Vα14 NKT cell line and conventional T cell line (B) were stimulated in vitro with Con A (10 μg/ml). 24 h later, the amounts of IL-4 or IFN-γ in the culture supernatants were measured by ELISA. Mean values in triplicate samples with standard deviations are shown. n.d., not detectable.

Article Snippet: The amounts of IL-4 and IFN-γ in the culture supernatants were assayed using ELISA kits (EN-2601-80 and EN-2604-50; Endogen, Inc.) according to the manufacturer's protocol.

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay