Drosophila larva transitions from larval stage L1 to larval stage L3 by intermittent molting. Prior to pupariation, L3 larvae stop feeding and migrate to a pupariation site on the side of a vial. Wandering L3 larvae were bisected and inverted in PBS, and all tissues were fixed in 4% formaldehyde/PBS for 30 min, permeabelized in 0.2% Triton X-100/PBS for 4 hours, and blocked in 3% BSA/PBS for 1 hour. All antibody stainings were performed in 3% BSA/PBS, incubation of primary and secondary antibodies were O/N. PBS was used for all rinses and washes (3× each for primary and secondary antibody incubation steps). Antibodies used were as follows: Chicken anti-RFP 1:2,000 (Rockland, 600-901-379); Goat anti-GFP 1:3,000 (Rockland, 600-101-215); Donkey anti-Goat Alexa Fluor 488 (Life Technologies, A11055); Donkey anti-Chicken Cy3 (Jackson ImmunoResearch, 703-165-155); and Donkey anti-Mouse Alexa Fluor 594 (Life Technologies, A21203). All secondary antibodies were used at 1:500. Tissues were dissected off the cuticle and were mounted in SlowFade Gold mounting medium with DAPI (Life Technologies, S36938). See Kockel et al. (2016) (link) for a detailed protocol.
Slowfade gold mounting medium with dapi
SlowFade Gold mounting medium with DAPI is a reagent designed for use in fluorescence microscopy applications. It is formulated to reduce photobleaching of fluorescent dyes, including DAPI, which is a nuclear counterstain. The product is intended to be used as part of the sample preparation process for microscopy analysis.
Lab products found in correlation
3 protocols using slowfade gold mounting medium with dapi
Drosophila Larval Tissue Staining
Drosophila larva transitions from larval stage L1 to larval stage L3 by intermittent molting. Prior to pupariation, L3 larvae stop feeding and migrate to a pupariation site on the side of a vial. Wandering L3 larvae were bisected and inverted in PBS, and all tissues were fixed in 4% formaldehyde/PBS for 30 min, permeabelized in 0.2% Triton X-100/PBS for 4 hours, and blocked in 3% BSA/PBS for 1 hour. All antibody stainings were performed in 3% BSA/PBS, incubation of primary and secondary antibodies were O/N. PBS was used for all rinses and washes (3× each for primary and secondary antibody incubation steps). Antibodies used were as follows: Chicken anti-RFP 1:2,000 (Rockland, 600-901-379); Goat anti-GFP 1:3,000 (Rockland, 600-101-215); Donkey anti-Goat Alexa Fluor 488 (Life Technologies, A11055); Donkey anti-Chicken Cy3 (Jackson ImmunoResearch, 703-165-155); and Donkey anti-Mouse Alexa Fluor 594 (Life Technologies, A21203). All secondary antibodies were used at 1:500. Tissues were dissected off the cuticle and were mounted in SlowFade Gold mounting medium with DAPI (Life Technologies, S36938). See Kockel et al. (2016) (link) for a detailed protocol.
Immunostaining of Drosophila Larval Neurons
Multicolor Immunofluorescence Staining
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