Peptide-specific Ab responses in mice were analyzed by ELISA as described. ELISA plates (Nunc MaxiSorp, Roskilde, Denmark) were coated with 2xP (5 μg/ml), blocked with 1% (w/v) BSA in PBST for 2 h, and incubated with mouse sera diluted 1:10 for IgE and 1:500 for IgG1 measurements. Bound IgE Abs were detected with an HRP-conjugated goat anti-mouse IgE antiserum (SouthernBiotech, Birmingham, AL) diluted 1:6000. Bound IgG Abs were detected with rat anti-mouse IgG1 mAb (BD Pharmingen, San Diego, CA) diluted 1:1000, followed by an HRP-conjugated goat anti-rat antiserum (Amersham Biosciences, Buckinghamshire, U.K.) diluted 1:2000. The specificity of the goat anti-mouse IgE antiserum for mouse IgE was confirmed by ELISA, as it detected mouse monoclonal IgE (28 (link)) but not mouse monoclonal IgG1 Abs (31 (link)) (data not shown). All values were calculated by subtraction of the appropriate buffer controls (22 (link), 32 (link)).
Hrp conjugated goat anti mouse ige antiserum
Manufactured by Southern Biotech
HRP-conjugated goat anti-mouse IgE antiserum is a laboratory reagent used to detect and quantify mouse IgE antibodies in various immunoassays. It is a polyclonal antibody generated in goats and conjugated to the enzyme horseradish peroxidase (HRP), which enables colorimetric or chemiluminescent detection.
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2 protocols using hrp conjugated goat anti mouse ige antiserum
1
ELISA for Peptide-specific Antibodies
2
ELISA for Peptide-specific Antibodies
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Peptide-specific Ab responses in mice were analyzed by ELISA as described. ELISA plates (Nunc MaxiSorp, Roskilde, Denmark) were coated with 2xP (5 μg/ml), blocked with 1% (w/v) BSA in PBST for 2 h, and incubated with mouse sera diluted 1:10 for IgE and 1:500 for IgG1 measurements. Bound IgE Abs were detected with an HRP-conjugated goat anti-mouse IgE antiserum (SouthernBiotech, Birmingham, AL) diluted 1:6000. Bound IgG Abs were detected with rat anti-mouse IgG1 mAb (BD Pharmingen, San Diego, CA) diluted 1:1000, followed by an HRP-conjugated goat anti-rat antiserum (Amersham Biosciences, Buckinghamshire, U.K.) diluted 1:2000. The specificity of the goat anti-mouse IgE antiserum for mouse IgE was confirmed by ELISA, as it detected mouse monoclonal IgE (28 (link)) but not mouse monoclonal IgG1 Abs (31 (link)) (data not shown). All values were calculated by subtraction of the appropriate buffer controls (22 (link), 32 (link)).
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