Plates were coated with 5µg/ml gag-p24 protein overnight. Following washing, plates were blocked with RPMI media containing 10% FBS (Gibco), 1% non essential amino acids (Gibco), 1% anti-anti cocktail (Gibco), and 0.1% 2- mercaptoethanol (Gibco). Cells were plated in a concentration of 1-2X 106 cells per well (GI) in triplicate and cultured for 24 hours at 37°C, washed, and isotype specific antibody was detected using HRP-conjugated goat anti-mouse IgA secondary (Southern Biotech). Spots were developed using BD ELISPOT AEC substrate set (551951) according to manufacturers protocol. Spots were quantified using a CTL-ImmunoSpot Analyzer and Software.
Hrp conjugated goat anti mouse iga secondary
Manufactured by Southern Biotech
The HRP-conjugated goat anti-mouse IgA secondary is a laboratory reagent designed for use in immunoassay techniques. It consists of goat-derived antibodies specific to mouse IgA immunoglobulins, conjugated to the enzyme horseradish peroxidase (HRP). This reagent can be used to detect and quantify the presence of mouse IgA in samples during analytical procedures.
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3 protocols using hrp conjugated goat anti mouse iga secondary
1
Gag-p24 Protein ELISA Assay
2
Gag-p24 Protein ELISA Assay
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Plates were coated with 5µg/ml gag-p24 protein overnight. Following washing, plates were blocked with RPMI media containing 10% FBS (Gibco), 1% non essential amino acids (Gibco), 1% anti-anti cocktail (Gibco), and 0.1% 2- mercaptoethanol (Gibco). Cells were plated in a concentration of 1-2X 106 cells per well (GI) in triplicate and cultured for 24 hours at 37°C, washed, and isotype specific antibody was detected using HRP-conjugated goat anti-mouse IgA secondary (Southern Biotech). Spots were developed using BD ELISPOT AEC substrate set (551951) according to manufacturers protocol. Spots were quantified using a CTL-ImmunoSpot Analyzer and Software.
3
B Cell Antibody Production Assay
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Plates were coated with 10 µg/ml CT (Sigma-Aldrich) overnight. After washing, plates were blocked with B cell media (RPMI containing 10% FBS [Gibco], 1% MEM NEAA [Gibco], 1% anti-anti [Gibco], and 0.1% 2-mercaptoethanol [Gibco]). Cells were plated in a concentration of 1–2 × 106 cells per well (GI) and cultured in B cell media for 24 h at 37°C, washed, and then isotype-specific antibody was detected using HRP-conjugated goat anti–mouse IgA secondary (Southern Biotech). Spots were developed using BD ELISPOT AEC substrate set (551951) according to manufacturer’s protocol. Spots were quantified using a CTL-ImmunoSpot Analyzer and Software.
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