Embryos (1–2 min post-wounding) were fixed in formaldehyde saturated heptane for 40 min. The vitelline membrane was removed by hand and the embryos were then washed 3 times with PAT [1x PBS, 0.1% Tween-20, 1% bovine serum albumin (BSA), 0.05% azide], then blocked in PAT for 2h at 4°C. Embryos were incubated with mouse anti-chickadee antibody (chi 1J; 1:10; Developmental Studies Hybridoma Bank) and guinea pig anti-Girdin antibody (1:500; provided by Patrick Laprise) [104 (link)] and for 24h at 4°C. Embryos were then washed 3 times with XNS (1x PBS, 0.1% Tween-20, 0.1% BSA, 4% normal goat serum) for 40 min each. Embryos were incubated with Alexa Fluor 568- and Alexa Fluor 633- conjugated secondary antibodies (1:1000; Invitrogen) overnight at 4°C. Embryos were washed with PTW (1x PBS, 0.1% Tween- 20), incubated with Alexa Fluor 488-conjugated Phalloidin at 0.005 units/μl (Molecular Probes/Invitrogen, Rockford, IL) at room temperature for 1 h, washed with PTW, and then imaged.
Alexa fluor 568 and alexa fluor 633 conjugated secondary antibodies
Manufactured by Thermo Fisher Scientific
Alexa Fluor 568- and Alexa Fluor 633-conjugated secondary antibodies are fluorescent-labeled secondary antibodies. These antibodies are designed for use in immunodetection techniques, such as immunohistochemistry and Western blotting, to visualize target proteins.
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Lab products found in correlation
2 protocols using alexa fluor 568 and alexa fluor 633 conjugated secondary antibodies
1
Immunofluorescence Staining of Drosophila Embryos
2
Immunostaining of Embryonic Actin Cytoskeleton
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Embryos (1-2 min post-wounding) were fixed in formaldehyde saturated heptane for 40 min. The vitelline membrane was removed by hand and the embryos were then washed 3 times with PAT [1x PBS, 0.1% Tween-20, 1% bovine serum albumin (BSA), 0.05% azide], then blocked in PAT for 2h at 4°C. Embryos were incubated with mouse anti-chickadee antibody (chi 1J; 1:10; Developmental Studies Hybridoma Bank) and guinea pig anti-Girdin antibody (1:500; provided by Patrick Laprise) [90] (link) and for 24h at 4°C. Embryos were then washed 3 times with XNS (1x PBS, 0.1% Tween-20, 0.1% BSA, 4% normal goat serum) for 40 min each. Embryos were incubated with Alexa Fluor 568-and Alexa Fluor 633-conjugated secondary antibodies (1:1000; Invitrogen) overnight at 4°C. Embryos were washed with PTW (1x PBS, 0.1% Tween-20), incubated with Alexa Fluor 488-conjugated Phalloidin at 0.005 units/μl (Molecular Probes/Invitrogen, Rockford, IL) at room temperature for 1 h, washed with PTW, and then imaged.
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