Immunohistochemistry for IL-4 and IL-17 was performed on 4 µm-thick lung tissue sections. Antigen on slides was retrieved with 1 mM EDTA (Boster) and slides were pretreated with 3% H2O2 (Boster) to quench endogenous peroxidase activity. After being blocked with normal goat serum, slides were incubated with anti-IL-4 (CAT#bs-0581-R) and anti-IL-17 (CAT#bs-1183-R) monoclonal antibodies (both Bioss) diluted 1:100 overnight at 4°C and treated with biotin-conjugated goat anti-rabbit IgG antibodies (CAT#bs-0295G-R, Bioss) diluted 1:50 for 30 min at room temperature. Slides were then stained with horseradish peroxidase (HRP)-conjugated streptavidin for 30 min and immunoperoxidase staining was developed using a 3,3′diaminobenzidine (DAB) for 5 min. IL-4- and IL-17-positive cells were examined using a BX-51 microscope and optical density (OD) was measured using ImageProPlus 6.0 software.
Anti il 4
Manufactured by Bioss Antibodies
Sourced in China
Anti-IL-4 is a laboratory reagent used for the detection and quantification of interleukin-4 (IL-4) in biological samples. IL-4 is a cytokine that plays a crucial role in the regulation of immune responses. The Anti-IL-4 product enables researchers to measure IL-4 levels, which can provide valuable insights into various physiological and pathological processes.
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Lab products found in correlation
Anti tnf α, by Bioss Antibodies (3 mentions)
Anti il 1β, by Bioss Antibodies (2 mentions)
Anti il 10, by Bioss Antibodies (2 mentions)
Anti il 6, by Bioss Antibodies (2 mentions)
Rabbit anti inos, by Bioss Antibodies (2 mentions)
Anti arg1, by Bioss Antibodies (2 mentions)
Alexa fluor 488 conjugated goat anti rabbit igg, by Bioss Antibodies (2 mentions)
7266 fluorescence microscope, by Leica (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Boster Bio (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Mouse anti iba1, by Bioss Antibodies (1 mentions)
Anti ifn γ, by ABclonal (1 mentions)
Ecl plus western blotting detection reagents, by Merck Group (1 mentions)
Anti β actin, by Bioss Antibodies (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bicinchoninic acid assay kit, by Beyotime (1 mentions)
4 protocols using anti il 4
1
Immunohistochemical Analysis of IL-4 and IL-17
2
Immunohistochemical Analysis of Neuroinflammation
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The rats were killed by intraperitoneal injection (10% chloral hydrated, 1 ml/100 g) and 4% paraformaldehyde (PFA) was used to fix via cardiac perfusion. Coronal brain slices (15 μm thick) were prepared and used for staining. Slices were washed in PBS for 5 min, three times, then blotted in 5% goat serum for 1 h at room temperature. Slices were incubated with the primary antibodies overnight at 4°C. Primary antibodies used: mouse anti-Iba1 (1:200 Sigma American), mouse anti-GFAP (1:200 Sigma American), rabbit anti-iNOS, anti-Arg1, anti-IL-1β, anti-TNF-α, anti-IL-4, anti-IL-6, and anti-IL-10 (1:200; Bioss, Beijing, China). After having been washed in PBS three times, these slices were incubated with secondary antibodies at 37°C for 2 h. Secondary antibodies: Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200; Bioss, Beijing, China), Cy3-conjugated goat ant-mouse IgG (1:200; Bioss, Beijing, China). Then these slices were washed three times again and stained with DAPI (Biyuntian, China) for 5 min. Results were imaged using a 7266-fluorescence microscope (Leica, Japan).
3
Immunofluorescence Assay for Microglial Markers
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Cells were fixed with 4% paraformaldehyde for 10 min and then washed in phosphate-buffered saline (PBS). After being blocked with 5% bovine serum albumin containing 0.5% Triton X-100 for 1 h at 37°C, the microglia were incubated with the following primary antibodies at 4°C overnight: mouse anti-Iba1 (1:200, Bioss, Beijing, China), rabbit anti-iNOS, anti-Arg1, anti-IL-1β, anti-TNF-α, anti-IL-4, anti-IL-6 and anti-IL-10 (1:200, Bioss). After being washed in PBS three times, the cells were incubated with the following secondary antibodies at 37°C for 1 h: Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200, Bioss) or Alexa Fluor Cy3-conjugated goat anti-mouse IgG (1:200, Bioss). After being washed in PBS three times, the cells were incubated with 4′,6-diamidino-2-phenylindole (DAPI; Biyuntian, China) for 5 min. The cells were then washed in PBS and observed under a 7266-fluorescence microscope (Leica, Japan).
4
Gestational Immune Profile Analysis
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Total protein was extracted from spleen tissues isolated from five pregnant CBA/J mice from each group on day 14 of gestation using RIPA lysis buffer (Beyotime Institute of Biotechnology). Protein (30 µg/lane) was quantified using a bicinchoninic acid assay kit (Beyotime Institute of Biotechnology) and separated by 10% SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 10% skim milk at room temperature for 4 h. Subsequently, the membranes were incubated at 4°C overnight with the following primary antibodies: Anti-FoxP3 (cat. no. bs-0269R; 1:1,000; BIOSS), anti-TNF-α (cat. no. A0277; 1:1,000; ABclonal Biotech Co., Ltd.), anti-IFN-γ (cat. no. bs-0480R; 1:1,000; BIOSS), anti-IL-4 (bs-0581R; 1:1,000; BIOSS) and anti-β-actin (cat. no. AC026; 1:2,000; ABclonal Biotech Co., Ltd.). Subsequently, the membranes were incubated with a goat anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody (1:2,000; cat. no. bs-0295G-HRP; BIOSS). Protein bands were visualized using ECL Plus Western Blotting Detection reagents (EMD Millipore). Blots were performed in triplicate and protein expression was quantified using ImageJ 2× software (National Institutes of Health) with β-actin as the loading control.
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