Purified PDCoV S1 protein with an Fc-tag was subjected to 12% SDS-PAGE and then transferred to a nitrocellulose (NC) membrane. The NC membrane was blocked using 5% (w/v) nonfat dried milk in phosphate-buffered saline (PBS) at 37 °C for 1 h and then incubated with a rabbit monoclonal antibody (mAb) against the Fc tag (1:20,000) and a porcine polyclonal antibody (pAb) against PDCoV (1:500) at 37 °C for 1 h. After washing three times with PBS containing 0.05% Tween 20 (PBST), the membrane was incubated in PBS containing horseradish peroxidase (HRP)-conjugated goat anti-rabbit (1:10,000) (Sigma) or rabbit anti-pig IgG (1:10,000) (Sigma) at 37 °C for 1 h. Membranes were washed with PBST and incubated with substrate solution (PBST, 0.05% DAB and 0.006% H2O2).
Rabbit anti pig igg
Manufactured by Merck Group
Sourced in United States
Rabbit anti-pig IgG is an antibody reagent produced by immunizing rabbits with pig immunoglobulin G (IgG). This reagent is designed for use in various laboratory applications that require the detection or quantification of pig IgG.
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Lab products found in correlation
2 protocols using rabbit anti pig igg
1
Western Blot Analysis of PDCoV S1 Protein
2
Indirect Immunofluorescence Assay for PSV
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The indirect immunofluorescence assay (IFA) was conducted as previously described with slight modification [36 (link)]. Cell monolayers were inoculated with PSV at an MOI of 0.01 and incubated for 12 h. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and blocked with 2% bovine serum albumin. A mAb specific for PSV-VP1 diluted 1:200 or swine serum at a 1:100 dilution was used as the primary antibody. The cells were then incubated with FITC-conjugated goat anti-mouse IgG 1:500 dilution (Thermo Fisher Scientific, Waltham, MA USA) or rabbit anti-pig IgG 1:50 dilution (Sigma, St. Louis, MO, USA). Nuclei were stained with 10 µg/mL DAPI (Solarbio, Beijing, China), and cells were then visualized under an inverted fluorescence microscope (Evos FL, Thermo-Fisher, Bothell, WA, USA).
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