Donkey anti goat igg fitc

The following primary antibodies were used for experiments: CD45-PE Cy7, CD45-APC Cy7, and CD45 APC (clone 30-F11, BD Biosciences, BioLegend), pan-keratin (Polyclonal, Dako), EpCAM-PE (clone G8.8, eBioscience), FoxN1 (clone G-20, Santa Cruz Biotechnology), FSP1 (clone S100A4, BioLegend), Thy1.2 (clone 30-H12, BD Biosciences), CD11b (clone, BioLegend), CD11b PerCpcy5.5 (clone M1/70, BioLegend), CD4-biotinylated (clone RM4-5, BioLegend), CD8-biotinylated (clone 53-6.7, BioLegend), CD19 (clone 1D3, BioLegend), rabbit anti-GFP (Life), and CD205 (LY75/DEC-205) (clone HD30 (Millipore). We also used lineage depletion panel containing B220. As control for EpCAM staining, we used rat IgG2a, κ isotype control (eBioscience). As control for CD45 staining, we used rat IgG2b, κ isotype control (BioLegend).
The following secondary reagents were used for experiments: donkey anti-rabbit IgG-TRITC, donkey anti-rabbit IgG-Cy5, donkey anti-rabbit IgG-FITC, donkey anti-rat IgG-TRITC, donkey anti-goat IgG-FITC, and goat anti-rat IgM-TRITC (Jackson ImmunoResearch); anti-rat IgG2a-FITC, anti-rat IgM-FITC, streptavidin-APC, streptavidin-APC Cy7, and streptavidin-PerCP Cy5.5 (BD Biosciences); and streptavidin-TRITC (Southern Biotechnology Associates).

The small intestine of the infected and euthanatized infant mice at 48 h post infection (hpi) were collected and checked histopathological changes. The small intestine sections were fixed in 10% (v/v) formalin in PBS for at least 48 h, then dehydrated in 70% (v/v) graded ethanol series and embedded in paraffin before being sectioned at the 4.0 μm thickness for further hematoxylin and eosin (H&E) staining. Histopathological observation of the small intestine was carried out under a light microscope. The jejunum paraffin sections of infected mice were assayed using Goat anti-rotavirus antibody (1:1000, made by our lab) and mouse anti-keratin 18 (red fluorescence represents keratin 18, #4548, Cell signaling, MA, USA). After incubation, slides were washed with PBS and incubated with donkey anti-Goat IgG-FITC (0.5 μg/mL, Jackson ImmunoResearch, West Grove, PA, USA) and rabbit anti-mouse IgG-DyLight™ 594 (0.5 μg/mL, Jackson ImmunoResearch, West Grove, PA, USA) for 1 h at 37 °C. The slides were then washed three times with PBS and covered with a cover slip, followed by staining with 4, 6-diamidino-2- phenylindole dihydrochloride (Sigma, USA). Samples were analyzed using a fluorescence microscope (Nikon, Tokyo, Japan).