A FluoroNunc Maxisorp 96-well plate was coated with 1 µg/mL 9D11 in PBS and incubated overnight at 4 °C. Wells were blocked with 200 µL TBS containing 1 mg/ml I (Human Albumin CSL 109697) for 1 hour at RT and washed thrice with TBS/Tw (TBS containing 0.05% v/v Tween-20 (8.17072.1000, Merck)). Samples and standard (purified C11 were diluted in TBS/Tw containing heat agg. nhIg 100 µg/ml (Octapharma 478393), incubated 63 °C 30 min., and subsequently added in duplicates. The plate was incubated at 4 °C overnight. Wells were washed thrice in TBS/Tw and incubated with biotinylated goat-anti-mouse IgG2b (1093-08, Southern Biotech), or biotinylated goat-anti-mouse IgG2c (1077-08, Southern Biotech) or biotinylated mouse-anti-mouse IgM(b) (553519 BD), 1 µg/ml TBS/Tw, at RT for 2 hours. Wells were washed 3 times in TBS/Tw, and Eu3 + -labeled streptavidin (1244-360, PerkinElmer) diluted 1:1,000 in TBS/Tw containing 25 µM EDTA was subsequently added to the wells and incubated at RT for 1 hour. Finally, the wells were washed thrice with TBS/Tw, and 200 µL enhancement buffer (AMPQ99800, Amplicon) was added. The plate was shaken for 5 minutes and counts were read in a time-resolved fluorometry plate reader (Victor X5, PerkinElmer).
Biotinylated goat anti mouse igg2b
Manufactured by Southern Biotech
Biotinylated goat-anti-mouse IgG2b is a secondary antibody that specifically binds to the IgG2b subclass of mouse immunoglobulins. It is conjugated with biotin, which allows for detection and amplification of signal in various immunoassay techniques.
Automatically generated - may contain errors
2 protocols using biotinylated goat anti mouse igg2b
1
Quantification of mouse immunoglobulin isotypes
2
Quantification of mouse immunoglobulin isotypes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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A FluoroNunc Maxisorp 96-well plate was coated with 1 µg/mL 9D11 in PBS and incubated overnight at 4 °C. Wells were blocked with 200 µL TBS containing 1 mg/ml I (Human Albumin CSL 109697) for 1 hour at RT and washed thrice with TBS/Tw (TBS containing 0.05% v/v Tween-20 (8.17072.1000, Merck)). Samples and standard (purified C11 were diluted in TBS/Tw containing heat agg. nhIg 100 µg/ml (Octapharma 478393), incubated 63 °C 30 min., and subsequently added in duplicates. The plate was incubated at 4 °C overnight. Wells were washed thrice in TBS/Tw and incubated with biotinylated goat-anti-mouse IgG2b (1093-08, Southern Biotech), or biotinylated goat-anti-mouse IgG2c (1077-08, Southern Biotech) or biotinylated mouse-anti-mouse IgM(b) (553519 BD), 1 µg/ml TBS/Tw, at RT for 2 hours. Wells were washed 3 times in TBS/Tw, and Eu3 + -labeled streptavidin (1244-360, PerkinElmer) diluted 1:1,000 in TBS/Tw containing 25 µM EDTA was subsequently added to the wells and incubated at RT for 1 hour. Finally, the wells were washed thrice with TBS/Tw, and 200 µL enhancement buffer (AMPQ99800, Amplicon) was added. The plate was shaken for 5 minutes and counts were read in a time-resolved fluorometry plate reader (Victor X5, PerkinElmer).
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