Estimation of IgM rheumatoid factors by fluorimetry.

Abstract

Stock borate saline buffer (Kotoulas and Moroz' 1971) containing 0O15 M sodium chloride and O-15 M boric acid was adjusted to pH 8 6 with sodium hydroxide and used as a 10% solution. Stock potassium phosphate buffer pH 7, 1 mol/l containing sodium azide (2 g/l) was diluted to 0-1 mol/l for use and Tween 20 (0 05%) was added to reduce nonspecific binding effects (Engvall and Perlmann, 1972). A stock solution (10 mg/ml) of rabbit FIl in phosphate buffer was prepared and filtered for use. It was stored at -20°C. Alkaline sodium dodecyl sulphate (SDS) was prepared using 2 g/100 ml 0 -1 N NaOH. MethodUsing a 1 cm diameter quartz cell in a Locarte digital fluorimeter with 13 stage PM tube and zinc lamp, the optimum sensitivity was achieved with a primary filter combination 805 Q (Ilford) and BG 12/2 mm (Schott) and secondary filters 805 Q, OG 515/2 mm, and Wratten 55 (Kodak). The instrument response was linear, turbidity interference minimal, and stability within experiments and between days high after a few minutes' warm-up. After treatment with ethanol and hot 40 % caustic soda solution, the plastic discs were rinsed 247 on 6 April 2019 by guest. Protected by copyright.

Incidence and specificity of antibodies to types I, II, III, IV, and V collagen in rheumatoid arthritis and other rheumatic diseases as measured by 125i‐radioimmunoassay

“…Hemagglutination of collagen-coated red blood cells, the most commonly reported method ( 3 3 , can result from the interaction of fibronectin (9,22) or glycosaminoglycans (8) with collagen and give false positive results. Solid phase radioimmunoassay (4,23) or fluorimetric assay (24,25) using immobilized collagen could give falsely high values due to binding of rheumatoid factor to small amounts of nonspecifically adsorbed IgG. Double antibody radioimmunoassays could detect labeled collagen nonspecifically adsorbed to immune complexes (9,26).…”

“…Twice the highest dilution of antibody giving 100% precipitation was routinely used. After incubation with second antibody for 18 hours at 4"C, precipitates were recovered by centrifugation at 3,OOOg (20) NSAlDs (24) * NSAIDs = nonsteroidal antiinflammatory drugs. Replicate samples never varied more than 10% and duplicate sera within a run also vaned by less than 10%.…”

Patterns of antihistone antibody specificity in systemic rheumatic disease. I. Systemic lupus erythematosus, mixed connective tissue disease, primary sicca syndrome, and rheumatoid arthritis with vasculitis

“…After incubation for 5 hours and then washing 3 times with Tween buffer, bound conjugate was released from the cuvette surface by addition of 750 pl of 0.2% sodium dodecyl sulfate (SDS) in 0.1M NaOH. The fluorescence of this solution was measured in a Locarte fluorimeter (13), using an SDS-NaOH solution blank. The fluorimeter was adjusted to read 1.000 with a 200-fold dilution of the mixture containing labeled antiserum in the SDS-NaOH solution.…”

Quantitative lasernephelometrische Bestimmung von Rheumafaktoren mittels lgG-beschichteter Latexpartikel

“…Der quantitamüngjöbulinklasse. Obwohl sie auch bei den anderen tive Nachweis ist mittels radioimmunologischer (10)(11)(12)(13)(14), sogenannten Kollagenosen und einigen weiteren chrofluoreszenzoptischer (15)(16)(17) oder enzymimmunologinisch entzündlichen Erkrankungen auftreten, wird Ihnen scher (18,19) Techniken möglich, die in der Regel sehr eine diagnostische Wertigkeit zuerkannt. Der Rheumaaufwendig und eher wissenschaftlichen Fragestellungen faktor-Test ist eines der diagnostischen Kriterien der vorbehalten sind.…”

“…Der Rheumaaufwendig und eher wissenschaftlichen Fragestellungen faktor-Test ist eines der diagnostischen Kriterien der vorbehalten sind. Lasernephelometrische Verfahren sind Arthritis and Rheumatism Association von 1958 (1) weniger aufwendig und liefern ebenfalls quantitative Erund 1963 (2) und auch in den New Yorker Kriterien für ge bnisse (9,15,20,21). Durch Präzipitation hitzeaggrerheumatoide Arthritis von 1966 enthalten.…”