K14.E7 mice and non-transgenic littermates were bled at 14-16 weeks of age. Sera were collected and stored at −20°C. Total serum IgE was measured by ELISA as described 72 (link). Briefly, An ELISA plate (Nunc MaxiSorp) was coated with purified anti-mouse IgE (BD Pharmingen clone R35-72, 1.25 μg/ml, 100 μl/well) in carbonate buffer pH9.2 at 4°C. Wells were washed and blocked with 200 μl of PBS 10% FCS. Purified mouse IgE isotype control (BD Pharmingen C38-2, 0.5mg/ml) antibody was used to create a standard curve, starting at 1000ng/ml. 100 μl/well of sera was incubated for 2h at room temperature. Biotin rat anti-mouse IgE was used as a detection antibody (BD Pharmingen clone R35-118, 1.25 μg/ml, 100 μl/well) and detected using a BD OptiEA Streptavidin HRP (BD Biosciences cat. 51-9002813, 100μl/well), and BD TMB substrate (BD Biosciences cat. 555214, 100μl/well). The plate was read at 450nm using a Synergy HT multi-mode plate reader (BioTek).
Optiea streptavidin hrp
Manufactured by BD
The BD OptiEA Streptavidin HRP is a laboratory reagent used in various immunoassay techniques. It consists of streptavidin, a protein that binds to biotin, conjugated with horseradish peroxidase (HRP), an enzyme that can catalyze a colorimetric reaction. This product is designed to facilitate the detection and quantification of biotinylated biomolecules in biological samples.
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Lab products found in correlation
2 protocols using optiea streptavidin hrp
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Measuring Serum IgE Levels in Mice
2
Measuring Serum IgE Levels in Mice
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K14.E7 mice and non-transgenic littermates were bled at 14-16 weeks of age. Sera were collected and stored at −20°C. Total serum IgE was measured by ELISA as described 72 (link). Briefly, An ELISA plate (Nunc MaxiSorp) was coated with purified anti-mouse IgE (BD Pharmingen clone R35-72, 1.25 μg/ml, 100 μl/well) in carbonate buffer pH9.2 at 4°C. Wells were washed and blocked with 200 μl of PBS 10% FCS. Purified mouse IgE isotype control (BD Pharmingen C38-2, 0.5mg/ml) antibody was used to create a standard curve, starting at 1000ng/ml. 100 μl/well of sera was incubated for 2h at room temperature. Biotin rat anti-mouse IgE was used as a detection antibody (BD Pharmingen clone R35-118, 1.25 μg/ml, 100 μl/well) and detected using a BD OptiEA Streptavidin HRP (BD Biosciences cat. 51-9002813, 100μl/well), and BD TMB substrate (BD Biosciences cat. 555214, 100μl/well). The plate was read at 450nm using a Synergy HT multi-mode plate reader (BioTek).
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