Specimens, either before gelation or after autoclave or LysC treatment, were incubated in 1× PBS with 0.1% Triton X-100 and 2% normal donkey serum (PBT) at room temperature (RT) for 2 hours for blocking, and in the case of pre-gelation specimens, permeabilization. Specimens were incubated with primary antibodies at 3 μg/mL in PBT, for 4 hours (RT), and then washed four times 30 minutes with PBT. Specimens were incubated with secondary antibodies at 20 μg/mL in PBT, for 4 hours (RT), and then washed four times at least 30 minutes with PBT. Secondary antibodies used were: goat Anti-Chicken Alexa 488 (Life Technologies), goat Anti-Rabbit Alexa 546 (Life Technologies) and goat Anti-Mouse CF633 (Biotium), except that goat Anti-Chicken Alexa 546 (Life Technologies) was used for Supplementary Fig. 1e, g(ii), h(ii) and goat Anti-Rabbit Alexa 488 (Life Technologies) was used for Fig. 1e .
Goat anti chicken alexa 546
Manufactured by Thermo Fisher Scientific
Goat Anti-Chicken Alexa 546 is a secondary antibody used in immunofluorescence applications. It is designed to detect and visualize chicken primary antibodies by binding to their Fc region and conjugating the Alexa Fluor 546 fluorescent dye, which emits red fluorescence when excited.
Automatically generated - may contain errors
2 protocols using goat anti chicken alexa 546
1
Immunofluorescence Staining Protocol
2
Immunofluorescence Staining Protocol
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Specimens, either before gelation or after autoclave or LysC treatment, were incubated in 1× PBS with 0.1% Triton X-100 and 2% normal donkey serum (PBT) at room temperature (RT) for 2 hours for blocking, and in the case of pre-gelation specimens, permeabilization. Specimens were incubated with primary antibodies at 3 μg/mL in PBT, for 4 hours (RT), and then washed four times 30 minutes with PBT. Specimens were incubated with secondary antibodies at 20 μg/mL in PBT, for 4 hours (RT), and then washed four times at least 30 minutes with PBT. Secondary antibodies used were: goat Anti-Chicken Alexa 488 (Life Technologies), goat Anti-Rabbit Alexa 546 (Life Technologies) and goat Anti-Mouse CF633 (Biotium), except that goat Anti-Chicken Alexa 546 (Life Technologies) was used for Supplementary Fig. 1e, g(ii), h(ii) and goat Anti-Rabbit Alexa 488 (Life Technologies) was used for Fig. 1e .
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