10–30 μl of fixed tongue dorsum scrapings were adhered to silanized slides as above, washed in 1× phosphate buffered saline with 0.1% Tween20 (PBST), then blocked with 10% goat serum in PBST for 1 h at room temperature in a humid chamber. Block was removed and anti-pan-cytokeratin (1:100, Santa Cruz Biotechnology #sc-81714) in 10% goat serum in PBST was applied to samples. A coverslip was applied to prevent evaporation and the slide was incubated in a humid chamber overnight at 4°C. After three PBST washes, samples were then incubated in AlexaFluor 647-conjugated goat anti-mouse IgG1 secondary antibody (1:500, Invitrogen #A-21240) in 10% goat serum in PBST for 1 h at room temperature in a humid chamber, washed with PBST, and cured overnight in the dark in Prolong Gold antifade mounting medium (Thermo Scientific).
Alexafluor 647 conjugated goat anti mouse igg1 secondary antibody
Manufactured by Thermo Fisher Scientific
AlexaFluor 647-conjugated goat anti-mouse IgG1 secondary antibody is a laboratory reagent used for detecting and visualizing mouse IgG1 primary antibodies in various immunological applications such as Western blotting, immunohistochemistry, and flow cytometry. The antibody is conjugated with the fluorescent dye AlexaFluor 647, which can be excited at 650 nm and emits light at 665 nm.
Automatically generated - may contain errors
2 protocols using alexafluor 647 conjugated goat anti mouse igg1 secondary antibody
1
Immunofluorescent Detection of Cytokeratins
2
Immunofluorescent Detection of Cytokeratins
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10–30 μl of fixed tongue dorsum scrapings were adhered to silanized slides as above, washed in 1× phosphate buffered saline with 0.1% Tween20 (PBST), then blocked with 10% goat serum in PBST for 1 h at room temperature in a humid chamber. Block was removed and anti-pan-cytokeratin (1:100, Santa Cruz Biotechnology #sc-81714) in 10% goat serum in PBST was applied to samples. A coverslip was applied to prevent evaporation and the slide was incubated in a humid chamber overnight at 4°C. After three PBST washes, samples were then incubated in AlexaFluor 647-conjugated goat anti-mouse IgG1 secondary antibody (1:500, Invitrogen #A-21240) in 10% goat serum in PBST for 1 h at room temperature in a humid chamber, washed with PBST, and cured overnight in the dark in Prolong Gold antifade mounting medium (Thermo Scientific).
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