Anti-Flag (A00187–200), anti-HA (A01244–100), anti-Actin (A00730–100), and protein G resin (L00209) were purchased from GenScript. Anti-NLRP3 (AG-20B-0014-C100) was purchased from Adipogen. Anti-GSDMD (ab209845) and anti-NEK7 (ab133514) were purchased from Abcam. Anti-ASC (67824) was purchased from CST. Anti-mouse caspase-1 was a kind gift from Dr Gabriel Núñez (University of Michigan Medical School). Ultra-pure LPS (tlrl-pb5lps), Nano-SiO2 (tlrl-sio), and poly(dA:dT)/lyovec (tlrl-patc) were purchased from InvivoGen. Nigericin (481990), doxycycline hydrochloride (10592–13–9), and EDTA-free protease inhibitor cocktail (11873580001) were purchased from Sigma. Salmonella strain SL1344 was originally from Dr Denise Monack (Stanford University). The constructs for NLRP3-Flag, NLRP3-SFP, and NEK7-HA have been previously described (17 (link)).
Anti gsdmd ab209845
Manufactured by Abcam
Sourced in United Kingdom
Anti-GSDMD (ab209845) is a primary antibody that can be used to detect the Gasdermin D (GSDMD) protein. GSDMD is a key mediator of pyroptosis, a form of inflammatory cell death. This antibody can be used in various applications, such as Western blotting and immunohistochemistry, to study the expression and localization of GSDMD in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti nek7 ab133514, by Abcam (2 mentions)
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Doxycycline hydrochloride, by Merck Group (1 mentions)
Anti flag, by GenScript (1 mentions)
Anti nlrp3 ag 20b 0014 c100, by Adipogen (1 mentions)
Protein g resin, by GenScript (1 mentions)
Edta free protease inhibitor cocktail, by Merck Group (1 mentions)
Enhanced chemiluminescence detection kit, by GE Healthcare (1 mentions)
Horseradish peroxidase, by Santa Cruz Biotechnology (1 mentions)
Il 1β, by Cell Signaling Technology (1 mentions)
β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)
Anti mouse caspase 1 ag 20b 0042, by Adipogen (1 mentions)
Anti β actin 66009 1 ig, by Proteintech (1 mentions)
Anti nlrp3 ag 20b 0014, by Adipogen (1 mentions)
Mitotracker, by Beyotime (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Protein a g agarose, by Beyotime (1 mentions)
Nigericin, by Merck Group (1 mentions)
Ab133514, by Abcam (1 mentions)
Poly da dt lyovec, by InvivoGen (1 mentions)
6 protocols using anti gsdmd ab209845
1
NLRP3 Inflammasome Activation Assay
2
Western Blotting of Inflammatory Markers
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Homogenized lung tissues were lysed in ice-cold radioimmunoprecipitation assay (Thermo Fisher Scientific, Waltham, MA) supplemented with protease inhibitor cocktail (Thermo Fisher Scientific), separated by electrophoresis on SDS-PAGE gels, and transferred to nitrocellulose transfer membranes (GE Amersham Biosciences, Pittsburgh, PA). Membranes were incubated with primary Abs overnight and then were incubated with corresponding secondary Abs conjugated to horseradish peroxidase (Santa Cruz Biotechnology). Signals were visualized using an enhanced chemiluminescence detection kit (GE Health). The Abs were used from different vendors as stated below: mouse Abs against p-NF-κB/p65 (sc-136548), caspase-1 (sc-514), TNF-α (sc52746), IL-6 (28343), and β-actin (sc-47778) were obtained from Santa Cruz Biotechnology (Dallas, TX). Rabbit Abs against NF-κB/p65 (no. 8242), STAT3 (no. 12640), p-STAT3 (no. 9145), and IL-1β (no. 12703) were obtained from Cell Signaling Technology (Danvers, MA). Rabbit Abs against GSDMD (anti-GSDMD, ab209845) were obtained from Abcam.
3
Investigating NLRP3 Inflammasome Activation
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Lipopolysaccharides (LPS, #L2880), adenosine triphosphate (ATP, #A3377), and aluminum potassium sulfate dodecahydrate (Alum, #237086) were purchased from Sigma (St. Louis, MO, USA). COS was bought from Abphyto Biotech Co., Ltd. (Chengdu, China). MitoSOX (#M36008) was obtained from Invitrogen (Carlsbad, CA, USA). MitoTracker (#C1049B) and protein A + G agarose (#P2012) were supplied by Beyotime (Shanghai, China). Anti-NLRP3 (#AG-20B-0014) and anti-mouse caspase-1 (#AG-20B-0042) antibodies were purchased from Adipogen (San Diego, CA, USA). Anti-NEK7 (#ab133514) and anti-GSDMD (#ab209845) antibodies were obtained from Abcam (Cambridge, UK). Anti-mouse IL-1β (#AF-401-NA) antibody was from R&D Systems (Minneapolis, MN, USA). Anti-Flag (#20543-1-AP), anti-HA (#66006-2-Ig), and anti-β-actin (#66009-1-Ig) antibodies were purchased from Proteintech (Rosemont, IL, USA).
4
Inflammasome Activation Pathway Analysis
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Anti-IL-1β antibodies (AF-401-NA) were from RD Systems, anti-GSDMD (ab209845) and anti-NEK7 antibodies (ab133514) were from Abcam, anti-caspase-1 (AG-20B-0042B) and anti-ASC antibodies (AG-25B-0006) were from Adipogen, and the anti-αtubulin antibodies (ARG65693) were from Arigo. LPS (Escherichia coli O111:B4; #L2630), ATP (A6419), and nigericin (481990) were obtained from Sigma. LND (AF-1890) and imiquimod (R-837) were obtained from Selleck. MSU crystals (tlrl-msu), muramyldipeptide (tlrl-mdp), flagellin from S. typhimurium (tlrl-stfla) and poly(dA:dT)/LyoVec™ were from Invivogen. Anti-hexokinase 2 antibodies (NBP2-02272) were from Novus. Anti-NLRP3 antibodies (15101) were from Cell Signaling Technology. Goat anti-rabbit IgG (HRP) (arg65351), donkey anti-goat IgG (HRP) (arg65352) and goat anti-mouse IgG antibodies (HRP) (arg65350) were from Arigo. Disuccinimidyl suberate (DSS) was purchased from Thermo Scientific.
5
Gut Microbiota and Metabolite Analysis
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Anti-ZO-1 (GB111981), anti-Occludin (GB111401), anti-F4/80 (GB11027), anti-IL-1β (GB11113), anti-MUC2 (GB11344), Oil Red staining solution (G1015), and PAS staining solution (G1008) were purchased from Servicebio. Anti-GSDMD (ab209845) was purchased from Abcam.
High-fat purified rodent diet (HF60-112,252) and control diet (AIN93G-110700) were obtained from Dyet. D-glucose (118,327,010) was procured from Guanghua Technology. Lipopolysaccharides (297–473-0), vancomycin (1404–93-9), neomycin sulfate (1405–10-3), metronidazole (443–48-1), ampicillin (69–52-3), acetone (67–64-1), DCA (D2510) and picryl sulfonic acid solution TNBS (P2297) were obtained from Sigma-Aldrich. Olive oil (O815211) was obtained from MACKLIN. Recombinant mouse GM-CSF (96–315-03–20) was procured from Peprotech. ATP solution (C0550) was purchased from Solarbio. L-012 (120–04,891) was acquired from Wako Science. Mouse DCA ELISA Kit (MM-44754M1) was bought from MEIMIAN-bio. Cholestyramine resin (11,041–12-6) was obtained from MedChem Express.
High-fat purified rodent diet (HF60-112,252) and control diet (AIN93G-110700) were obtained from Dyet. D-glucose (118,327,010) was procured from Guanghua Technology. Lipopolysaccharides (297–473-0), vancomycin (1404–93-9), neomycin sulfate (1405–10-3), metronidazole (443–48-1), ampicillin (69–52-3), acetone (67–64-1), DCA (D2510) and picryl sulfonic acid solution TNBS (P2297) were obtained from Sigma-Aldrich. Olive oil (O815211) was obtained from MACKLIN. Recombinant mouse GM-CSF (96–315-03–20) was procured from Peprotech. ATP solution (C0550) was purchased from Solarbio. L-012 (120–04,891) was acquired from Wako Science. Mouse DCA ELISA Kit (MM-44754M1) was bought from MEIMIAN-bio. Cholestyramine resin (11,041–12-6) was obtained from MedChem Express.
6
LPS, Nigericin, and DOTAP Modulation of Inflammatory Responses
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LPS (Escherichia coli O111:B4, L2630), nigericin (N7143) and N-(2,3-dioleoyloxy-1-propyl)trimethylammonium methyl sulfate (DOTAP) liposomal transfection reagent (144189-73-1) were from Sigma-Aldrich. Ultrapure flagellin from S. typhimurium (tlrl-epstfla-5) and poly(dA:dT; tlrl-patn) were purchased from InvivoGen. TcdB from List Labs (155). Lipofectamine RNAiMAX Transfection Reagent (13778150) was from Invitrogen. Mouse TNF-α (88-7324-88), IL-1β (88-7013-88), IL-18 (BMS618-3), and IL-6 (88-7064-22) ELISA kits were from Invitrogen. The LightShift Chemiluminescent EMSA Kit (20148) was from Thermo Fisher Scientific Inc. Anti–IL-1β (AF-401-NA) was from R&D Systems. Anti–caspase-1 (AG-20B-0042) and anti-ASC (AG-25B-0006-C100) were from AdipoGen. Anti-GSDMD (ab209845) and anti-H3K27ac (ab4729) were from Abcam. Antiactin (sc-47778), anti-IRF8 (sc-365042), anti-RNAPII (sc-47701), and anti-PU.1 (sc-390405) were from Santa Cruz Biotechnology, Inc. Anti-Brd4 (A301-985A) was from Bethyl Laboratories. Anti-H3K4me1 (07-436) and anti-H3K4me3 (05-745R) were from EMD Millipore.
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