Rabbit anti rfp 600 401 379

Third instar larvae were fixed and stained according to standard protocols. Briefly, larvae fillet preps were fixed in 4% paraformaldehyde/PBS for 20 min at room temperature, and stained with the proper primary antibodies and subsequent secondary antibodies, each for 2 h at room temperature. The following antibodies were used: chicken anti-GFP (ab13970, 1:100, Abcam), goat anti-HRP (123-605-021, 1:200, Jackson ImmunoResearch), rabbit anti-RFP (600-401-379, 1:500, Rockland Immunochemicals), rabbit anti-Ca-α1D (1:50)^{41 (link)}, rabbit anti-Ca-α1D II (1:100, this study), rabbit anti-Ca-β (1:100, this study), rabbit anti-Irk1 (1:100, this study), and fluorescence-conjugated secondary antibodies (1:1000, Jackson ImmunoResearch).

Tissues or cells were lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1 mM NaF, 1 mM sodium orthovanadate, and 1 mM phenylmethylsulfonyl fluoride. An equal amount of total protein (25 μg/lane for cells, 50 μg/lane for tissues) was loaded into 3–8% NuPAGE Tris-acetate gels (Invitrogen) and transferred to poly-vinylidene fluoride membranes for detection with indicated antibodies. The following antibodies were used: rabbit anti-RFP (600-401-379, Rockland) at 1:1,000 dilution; mouse anti-GADPH (60004-1-Ig, Proteintech) at 1:5,000 dilution.