The method for fluorescence microscopy of oocytes was adopted from Löbel et al.61 with minor modifications. Oocytes were injected with either water or 30 ng of cRNA with protein expression occurring over 2 days at 17°C. Oocytes were collected, washed twice with phosphate-buffer saline (PBS), then stained with 0.05 mg ml−1 CF633-conjugated wheat germ agglutinin (Biotium) in PBS for 5 min at room temperature. Oocytes were then washed with PBS. CF633 and eGFP fluorescence were measured using a Leica SP8 upright confocal microscope equipped with a 10× objective lens, using He-Ne (633 nm) and argon (488 nm) lasers, for CF633 and eGFP, respectively.
Cf633 conjugated wheat germ agglutinin
Manufactured by Biotium
CF633-conjugated wheat germ agglutinin is a fluorescently labeled lectin that binds to N-acetylglucosamine and sialic acid residues on cell surfaces. It can be used to label and visualize cell surface glycoconjugates.
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2 protocols using cf633 conjugated wheat germ agglutinin
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Fluorescence Microscopy of Oocytes
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Fluorescence Microscopy of Oocytes
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The method for fluorescence microscopy of oocytes was adopted from Löbel et al.59 with minor modifications. Oocytes were injected with either water or 30 ng of cRNA with protein expression occurring over 2 days at 17°C. Oocytes were harvested, washed twice with PBS, then stained with 0.05 mg ml−1 CF633-conjugated wheat germ agglutinin (Biotium) in PBS for 5 min at RT. Oocytes were then washed with PBS. CF633 and eGFP fluorescence were measured using a Leica SP8 upright confocal microscope equipped with a 10× objective lens, using He-Ne (633 nm) and Argon (488 nm) lasers, for CF633 and eGFP, respectively.
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