Cy5 conjugated anti mouse ig

Samples were fixed in 4% formaldehyde or 4% methanol-free formaldehyde in phosphate buffer saline (PBS, Fisher-Scientific, pH = 7.4; Cat No. #BP399-4). Tissues were permeabilized in PBT (PBS with 0.1–0.3% Triton X-100, pH = 7.4) and immunohistochemistry was performed using standard procedures (Ashburner, 1989 ). The following antibodies were provided by the Developmental Studies Hybridoma Bank (Iowa City, IA): mouse anti-Bruchpilot (Brp, 1:20), mouse anti-Neurotactin (BP106, 1:10), rat anti-DN-Cadherin (DN-EX #8, 1:20), and mouse anti-Neuroglian (BP104, 1:30). Secondary antibodies, IgG₁ (Jackson ImmunoResearch; Molecular Probes) were used at the following dilutions: Cy5-conjugated anti-rat Ig (1:100), Cy3-conjugated anti-mouse Ig (1:200), Cy5-conjugated anti-mouse Ig (1:250); Alexa 546-conjugated anti-mouse (1:500), DynaLight 649-conjugated anti-rat (1:400), Alexa 568-conjugated anti-mouse (1:500).

Samples were fixed in 4% formaldehyde or 4% methanol-free formaldehyde in phosphate buffer saline (PBS, Fisher-Scientific, pH = 7.4; Cat No. #BP399-4). Tissues were permeabilized in PBT (PBS with 0.1–0.3% Triton X-100, pH = 7.4) and immunohistochemistry was performed using standard procedures (Ashburner 1989 ). The following primary antibodies were provided by the Developmental Studies Hybridoma Bank (Iowa City, IA): mouse anti-bruchpilot (nc82; 1:20); rat anti-DN-Cadherin (DN-EX #8, 1:20), and mouse anti-Neurotactin (BP106, 1:10). Additional primary antibodies: rabbit anti-Deadpan (Dpn; Bier et al., 1992 (link)). Secondary antibodies, IgG₁ (Jackson ImmunoResearch; Molecular Probes) were used at the following dilutions: Cy5-conjugated anti-rat Ig (1:100), Cy3-conjugated anti-mouse Ig (1:200), Cy5-conjugated anti-mouse Ig (1:250); Alexa 546-conjugated anti-mouse (1:500), DynaLight 649-conjugated anti-rat (1:400), Alexa 568-conjugated anti-mouse (1:500).