Dye injection and immunohistochemistry methods have previously been described in detail (Boerner and Godenschwege, 2010 (link), 2011 ). In brief, the dissected animal CNS was mounted dorsal side up on VECTABOND™ (Vector Labs) coated 0.9-1.1 mm etched slides. A glass electrode (80-100 MΩ) filled with a dye solution of 10% w/v neurobiotin (Vector Labs) and tetramethyl rhodamine-labeled dextran (Invitrogen) backfilled with 2 M potassium acetate was used to inject the dyes into the GF axons by passing depolarizing current. Samples fixed in 4% paraformaldehyde were prepared for confocal microcopy as described previously (Boerner and Godenschwege, 2010 (link), 2011 ). The following antibodies were used: streptavidin-Cy2 conjugate (Jackson ImmunoResearch; 1:750), anti-GFP A11122 (Invitrogen, 1:500), goat anti-rabbit-Cy2 (Jackson ImmunoResearch, 1:500 dilution) to visualize neurobiotin or GFP. Samples were scanned at a resolution of 1024×1024 pixels, 2.5× zoom, and 0.5 μm step size with a Nikon C1si Fast Spectral Confocal system using a 60× oil immersion objective lens. Images were processed using Nikon Elements Advance Research 4.0 and Adobe Photosuite CS4 software.
Streptavidin cy2 conjugate
Manufactured by Jackson ImmunoResearch
Streptavidin-Cy2 conjugate is a laboratory reagent used for the detection and visualization of biotinylated molecules. Streptavidin, a bacterial protein, is conjugated to the fluorescent dye Cy2, enabling the labeling and detection of biotin-tagged targets.
Automatically generated - may contain errors
Lab products found in correlation
Vectabond, by Vector Laboratories (2 mentions)
Neurobiotin, by Vector Laboratories (2 mentions)
Tetramethyl rhodamine labeled dextran, by Thermo Fisher Scientific (2 mentions)
C1si fast spectral confocal system, by Nikon (2 mentions)
Cy2 goat anti rabbit, by Jackson ImmunoResearch (1 mentions)
Anti gfp a 11122, by Thermo Fisher Scientific (1 mentions)
2 protocols using streptavidin cy2 conjugate
1
Dye Injection and Immunohistochemistry for Neuronal Labeling
2
Dye Injection and Immunohistochemistry of Drosophila Nervous System
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Dye injection and immunohistochemistry methods have previously been described in detail62 (link),63 (link). In brief, the ventral nerve cord of adult Drosophila was dissected and mounted dorsal side up on VECTABOND™ (Vector Labs) coated 0.9–1.1 mm etched slides. An 80–100 MΩ glass electrode filled with a dye solution of 10% w/v neurobiotin (Vector Labs) and tetramethyl rhodamine-labeled dextran (Invitrogen) and backfilled with 2 M potassium acetate was used to inject the dyes into the GF axons by passing depolarizing current. Samples were fixed in 4% paraformaldehyde and were prepared for confocal microscopy as described previously62 (link),63 (link). Streptavidin-Cy2 conjugate (Jackson ImmunoResearch; 1:750) was used to visualize neurobiotin. Samples were scanned at a resolution of 1024 × 1024 pixels, 2.5× zoom, and 0.5 μm step size with a Nikon C1si Fast Spectral Confocal system using a 60× oil immersion objective lens. Dye filling of the GFs with Lucifer Yellow was performed as described in ref. 15 (link).
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