Testicular tissue sections were dewaxed and rehydrated, followed by Sodium Citrate Antigen Retrieval Solution (Solarbio, Beijing, China) at 95 °C for 15 min. Tissue permeabilization was conducted using phosphate buffered saline (PBS) containing Triton X-100. Tissue sections were blocked with 10% goat serum diluted in PBS at room temperature for 30 min. The VASA antibody (Gene Tex, Irvine, CA, USA) was diluted 1:500 and incubated at room temperature for 30 min, followed by incubation at 4 °C for 24 h, then returned to room temperature for an additional 30 min. Sections were washed five times with PBS for 3 min each. Goat Anti-Rabbit IgG/FITC (Solarbio, Beijing, China) was diluted 1:200 and incubated at room temperature for 1 h. This was followed by five PBS washes for 3 min each. DAPI staining (Solarbio, Beijing, China) solution was diluted 1:1000 and applied for 1 min, then sections were washed 3–5 times with PBS. Slides were mounted with 50% glycerol and observed and photographed under a fluorescence microscope.
Goat anti rabbit igg fitc
Manufactured by Solarbio
Sourced in China, United States
Goat Anti-Rabbit IgG/FITC is a secondary antibody used in immunological assays. It is produced by immunizing goats with rabbit immunoglobulin G (IgG) and conjugating the resulting antibodies with the fluorescent dye FITC. This product can be used to detect and visualize rabbit primary antibodies in various applications, such as immunofluorescence, flow cytometry, and Western blotting.
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3 protocols using goat anti rabbit igg fitc
1
Immunofluorescent Detection of VASA in Testicular Tissue
2
Immunofluorescence Analysis of PI3K/Akt/HSP70 Pathway
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Cells were cultured for 24 to 48 h before the MIRI model was established. Subsequently, the cells were washed three times with PBS and fixed with 4% paraformaldehyde (cat. no. P1110; Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 30 min, then blocked with 10% goat serum (cat. no. SL038; Beijing Solarbio Science & Technology Co., Ltd.) at room temperature for 30 min. The cells were incubated with the primary antibody overnight at 4˚C using PI3K (1:5,000; cat. no. 09-482; MilliporeSigma), phosphorylated (p)-Akt (1:200; cat. no. ab81283; Abcam), and HSP70 (1:50; cat. no. ab181606; Abcam). The cells were then washed and incubated with goat anti-mouse IgG-FITC (1:500; cat. no. SA0015; Beijing Solarbio Science & Technology Co., Ltd.) or goat anti-rabbit IgG-FITC (1:500; cat. no. SA0025; Beijing Solarbio Science & Technology Co., Ltd.) for 30 min without light at 37˚C. Nuclei were stained with a DAPI solution (cat. no. C0065; Beijing Solarbio Science & Technology Co., Ltd.) at 37˚C for 5 min. The cells were mounted with an anti-fluorescence attenuator and the staining was observed under a fluorescence microscope.
3
Identifying Isolated BVDV Strains
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To identify the isolated BVDV strains, monolayer MDBK cells were infected with the 5th passages of the isolated viruses. Mock-infected cells were used as the negative controls. After culturing for 48 h, the cells were fixed with 4% paraformaldehyde, incubated with 0.2% TritonX-100, and blocked with 1% bovine serum albumin. Then, the cells were incubated with BVDV monoclonal antibodies (VMRD, Washington, WA, USA) and Goat anti-Rabbit IgG/FITC (Solarbio®, Beijing, China) before examination using the fluorescence microscope.
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