Anti rat cy5

Primary antibodies: anti-CDK5RAP2 (this work), anti-Cep192 (this work), anti-CP55 (rat, this work), anti-CP148 [16 (link)], anti-CP91 [31 (link)], anti-CP224 [32 (link)], YL1/2 [33 (link)], and anti-NE81 [34 (link)].
Secondary antibodies: all AlexaFluor conjugates were purchased from Thermo Fisher Scientific (Darmstadt, Germany), anti-mouse-Cy5 and anti-rat-Cy5 from Jackson Labs (Dianova, Hamburg, Germany), and enzyme conjugates for Western blotting from Sigma (Deisenhofen, Germany). Strept488 und AP.

For analysis of radioactivity in harvested tissues, tissues were weighed, and radioactivity was quantified using a γ- counter (1282-Compugamma, LKB-Wallac) together with calibration standards. Data were expressed as %ID/g.
Skin grafts were harvested alongside normal mouse skin of similar size (from the lower back) for histology and frozen in Optimal Cutting Temperature (OCT) compound followed by tissue sectioning (8 μm). Thawed sections were fixed in 4% paraformaldehyde/PBS for 10 min, permeabilized (0.2% [v/v] Triton X-100/PBS), washed, and blocked (20% [v/v] goat serum/PBS containing 0.1% fish skin gelatin and 0.25% [v/v] Tween 20) before being stained with the following primary antibodies (2 μg/mL) overnight at 4°C: anti-human CD3 (polyclonal rabbit, A0452, Dako), anti-FOXP3 (monoclonal rat, clone PCH101, eBioscience), anti-Gr-1 (monoclonal rat, RB6-8C5, eBioscience), and anti-GFP (monoclonal mouse, 3E6, Invitrogen). After washing, sections were stained with secondary antibodies: goat anti-mouse-AlexaFluor 555, anti-rabbit-AlexaFluor 568 (Invitrogen), and anti-rat-Cy5 (Jackson ImmunoResearch). Nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI; 10 μg/mL PBS). Samples were mounted using fluorescence mounting medium (Dako) and imaged on a NikonA1 confocal microscope. Fiji/ImageJ v.1.5 software was used for analysis.