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Anti gsdmd

Manufactured by ABclonal
Sourced in China, United States

Anti-GSDMD is a primary antibody that specifically recognizes GSDMD (Gasdermin D), a protein involved in the process of pyroptosis, a form of programmed cell death. This antibody can be used for various applications, such as Western blotting, immunohistochemistry, and immunocytochemistry, to detect and study the expression and localization of GSDMD in biological samples.

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3 protocols using anti gsdmd

1

Comprehensive Western Blot Protocol for Immunological Markers

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The Western blot protocol used was previously described [7 (link)]. The primary antibodies included anti-TLR4 (Proteintech, China), anti-MyD88 (Proteintech), anti-NF-kB (R&D Systems, U.S.A.), anti-RIPK3 (Proteintech), anti-MLKL (Proteintech), anti-NLRP3 (ZEN BIO, China), anti-Caspase1 (Novus, U.S.A.), anti-Caspase8 (Proteintech), anti-GSDMD (ABclonal, China), anti-IL-1β (R&D Systems), and anti-β-actin (Elabscience, China). An anti–rabbit (or goat or mouse or rat) antibody (ABclonal) were applied to the secondary antibody.
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2

Western Blot Analysis of INS-1 Cell Proteins

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Total protein of INS-1 cells was extracted and lysed in RIPA lysis buffer (Beyotime, Shanghai, China) including the protease inhibitor (Roche, CA, USA). A BCA protein assay kit (Beyotime, Shanghai, China) was used to quantify proteins, and samples were mixed with 1 SDS sample buffer (50 mM Tris, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, and 0.01% bromophenol blue). Proteins were separated in 12% SDS-PAGE, transferred onto the PVDF membrane, and immunoblotted with anti-GSDMD (1 : 1000, ABclonal, USA) and anti-NLRP3 (1 : 1000, ABclonal, USA) at 4°C overnight. Secondary antibodies (goat anti-rabbit IgG H&L (HRP), Abcam, USA) were applied for 2-3 h at room temperature, and membranes were developed via the BCA protein colorimetric assay kit (Beyotime, Shanghai, China). Developed protein bands were quantified by Tanon Gel Image System V2.0 program (Shanghai, China).
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3

Immunofluorescence Analysis of JEV Infection

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Cells were seeded on coverslips, and when the cells grew to 90%, they were infected with JEV at MOI = 1, and a blank control group was set up. Then, 48 h after infection, the cells were fixed with 4% paraformaldehyde, and the cells were permeabilized and sealed by conventional methods after fixation. Anti-GSDMD (ABclonal, Wuhan, China) and anti-JEV E (homemade) primary antibodies (1:200) were then incubated overnight at 4 °C. After washing, cells were incubated with FITC-conjugated and CY3-conjugated secondary antibodies (1:500) (ABclonal) for 1 h at room temperature. Finally, nuclei were stained with Hoechst and visualized with a fluorescence microscope.
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