NPCTL- and M2e-specific antibodies were measured by ELISA. Briefly, Nunc-Immuno plates (Nunc 442404) were coated with 50 μL PBS-diluted M2e peptide (for M2e-immunized positive control) or NPCTL peptide (for all NPCTL-immunized samples), 5 μg/ml, at 4°C overnight. Unspecific binding was blocked with 200 μl of 0.25% gelatin in PBS-T solution (0.5% Tween-20 in PBS). Fifty microliters of the serially diluted sera was added to each well and incubated at 37°C for one and half hours. After extensive washing with PBS-T solution, binding antibodies were detected by horseradish peroxidase-labeled goat anti-mouse IgG antibody (Agilent Dako; P044701–2) and o-phenylenediamine dihydrochloride substrate (Sigma), following the manufacturer’s instructions.
Horseradish peroxidase labeled goat anti mouse igg antibody
Manufactured by Agilent Technologies
Horseradish peroxidase-labeled goat anti-mouse IgG antibody is a secondary antibody used in various immunoassay techniques. It binds to mouse primary antibodies and provides a means of detection through the enzymatic activity of the horseradish peroxidase label.
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2 protocols using horseradish peroxidase labeled goat anti mouse igg antibody
1
ELISA for M2e and NPCTL Antibodies
2
Measuring NPCTL and M2e Antibodies by ELISA
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NPCTL‐ and M2e‐specific antibodies were measured by ELISA. Briefly, Nunc‐Immuno plates (Nunc 442404) were coated with 50 μL PBS‐diluted M2e peptide (for M2e‐immunized positive control) or NPCTL peptide (for all NPCTL‐immunized samples), 5 μg/ml, at 4°C overnight. Unspecific binding was blocked with 200 μl of 0.25% gelatin in PBS‐T solution (0.5% Tween‐20 in PBS). Fifty microliters of the serially diluted sera was added to each well and incubated at 37°C for one and half hours. After extensive washing with PBS‐T solution, binding antibodies were detected by horseradish peroxidase‐labeled goat anti‐mouse IgG antibody (Agilent Dako; P044701‐2) and o‐phenylenediamine dihydrochloride substrate (Sigma), following the manufacturer's instructions.
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