Following CpG stimulation (5 days), we performed an ELISA to detect secreted immunoglobulin in the supernatant of the stimulated cells. Briefly, 96-well plates (Costar3590 EIA/RIA plate) were coated overnight with purified goat anti-human IgA+IgG+IgM (H+L chain; Jackson ImmunoResearch Laboratories). After washing with PBS containing 0.05% Tween and blocking with PBS containing 1% gelatin (1 h, room temperature), plates were incubated for 1 h at 37°C with the supernatants of cultured cells. After removing the supernatants and washing, plates were incubated for 1 h with peroxidase-conjugated fragment goat anti-human IgA or IgG or IgM antibodies (Jackson ImmunoResearch Laboratories). The assay was developed with O-phenylendiamine tablets (Sigma-Aldrich) as a chromogenic substrate. Absorbance at 450 nm was measured, and immunoglobulin concentrations were calculated by interpolation with the standard curve.
Peroxidase conjugated fragment goat anti human iga or igg or igm antibodies
Manufactured by Jackson ImmunoResearch
Peroxidase-conjugated fragment goat anti-human IgA or IgG or IgM antibodies are secondary antibodies used for the detection and quantification of human immunoglobulins. The antibodies are conjugated with the enzyme horseradish peroxidase, which catalyzes a colorimetric reaction that can be measured spectrophotometrically.
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Lab products found in correlation
2 protocols using peroxidase conjugated fragment goat anti human iga or igg or igm antibodies
1
ELISA for Secreted Immunoglobulin
2
ELISA Quantification of Secreted Immunoglobulins
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Secreted Igs were detected on day 7 by ELISA. Briefly, 96-well plates (Corning Inc., Corning, NY, USA) were coated overnight with purified goat anti-human IgA plus IgG, plus IgM (Jackson ImmunoResearch Laboratories). After washing with PBS containing 0.05% Tween and blocking with PBS containing 1% gelatin (1 h, room temperature), plates were incubated for 1 h at 37°C with the supernatants of the cultured cells. After washing, plates were incubated for 1 h with peroxidase-conjugated fragment goat anti-human IgA or IgG or IgM antibodies (Jackson ImmunoResearch Laboratories). The assay was developed with O-phenylendiamine tablets (Sigma-Aldrich) as a chromogenic substrate. Absorbance at 405 nm was measured, and Ig concentrations were calculated by interpolation with the standard curve [45 (link)].
Specific anti-PnPS IgG serum concentrations were measured using VaccZymeTM Anti-PCP IgG Enzyme Immunoassay kit from Binding Site (The Binding Site) according to the manufacturer's instructions. For anti-PCP, IgG concentrations were calculated from the standard curves in milligrams per liter.
Specific anti-PnPS IgG serum concentrations were measured using VaccZymeTM Anti-PCP IgG Enzyme Immunoassay kit from Binding Site (The Binding Site) according to the manufacturer's instructions. For anti-PCP, IgG concentrations were calculated from the standard curves in milligrams per liter.
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