Anti gfp 3e6

Third-instar larval preparations (muscles 6/7) were filleted and fixed in 4% paraformaldehyde, washed, and incubated overnight at 4°C with primary antibodies. Secondary antibodies were applied at room temperature for 2 hr. The following primary antibodies were used: anti-Myc (9E10 Santa Cruz), anti-GFP 3E6 (1:500; mouse; Life Technologies), anti-NC82 (1:100; mouse; Developmental Studies Hybridoma Bank), anti-HA antibody (1:1000; Rabbit; cell signaling technology) and anti-DLG (1:10,000; rabbit). Alexa-conjugated secondary (488, 555) antibodies and Cy5-conjugated goat ant-HRP were used at 1:500 (Life Technologies; Molecular Probes). Larval preparations were mounted in Vectashield (Vector) and imaged with an Axiovert 200 (Zeiss) inverted microscope, a 100X Plan Apochromat objective (1.4 NA) and a cooled charge-coupled device camera (Coolsnap HQ, Roper). Slidebook 5.0 Intelligent Imaging Innovations (3I) software was used to capture, process and analyze images. Structured illumination microscopy imaging was performed using the N-SIM Nikon system, consisting of a Nikon Ti-E Microscope equipped with a Apo TIRF 100×/1.49 Oil objective and an Andor DU897 Camera.