Incubation:Article Title: STING ‐dependent induction of neutrophilic asthma exacerbation in response to house dust mite
Article Snippet: Membranes 35 were blocked with 5% nonfat milk (Cell signaling, Massachusetts) in 1X TBS-T (20 mMTris 36 Base, 150 mM sodium chloride, and 0.05% Tween-20 pH 7.6) for 1h at room temperature. .. 37 Full membranes or portions of membranes were incubated overnight using primary antibodies 38 from rabbit anti-phospho-STAT6 (#56554 1/500; Cell signaling), anti-STAT6 (#5397 1/500; 39 1/500; Cell signaling), anti-γH2AX (#7631 1/500; Cell signaling), anti-phospho-STING 41 (#72971 1/500; Cell signaling), anti-STING (#13647 1/500; Cell signaling), anti-phospho-42 TBK1 (#5483 1/500; Cell signaling), anti-TBK1 (#3504 1/500; Cell signaling), anti-phospho-43 IRF3 (#4947 1/500; Cell signaling), IRF3 (#4302 1/500; Cell signaling), anti- cGAS (#31659 44 1/500; Cell signaling), anti-DDX41 (#15076 1/500; Cell signaling), anti-IFI204 (#ab228512 45 1/500; Abcam), anti-AIM2 (#63660 1/500; Cell signaling), anti-NLRP3 (#15101 1/500; Cell 46 signaling), anti-caspase-11/cleaved-caspase-11 (#NB120-10454 1/500; Novus), anti-IL-1β 47 (#12426 1/500; Cell signaling), anti-IL-18 (#57058 1/500; Cell signaling), anti-cleaved 48 caspase-3 (#9661 1/500), anti-caspase-3 (#9662 1/500; Cell signaling), anti-MLKL (#37705 49 1/500; Cell signaling), anti-cleaved GSDMD (#10137 1/500; Cell signaling), anti-GSDMD 50 (#ab219800 1/500; Abcam), mouse anti-ZBP1 (#SC-271483 1/300; Santa Cruz, Texas), anti-51 caspase-8 (#4927 1/500; Cell signaling), anti-RIPK3 (#95702, 1/500, Cell signaling), anti IL-52 25 (# PA5-100835, 1/500, Invitrogen) and anti-actin (#A3854 1/10000; Sigma-Aldrich, 53 Massachusetts). .. Membranes were washed in TBS-T three times for 10 min each at room 54 temperature, and then incubated with goat anti-rabbit-IgG-HRP-conjugate (#7074 1/2000; Cell 55 signaling) or horse anti-mouse-IgG-HRP-conjugate (#7076 1/2000; Cell signaling) diluted in 56 5% non-fat milk in TBS-T for 1h at RT.
Article Title: NINJ1 mediates inflammatory cell death, PANoptosis, and lethality during infection conditions and heat stress.
Article Snippet: .. Following electrophoretic transfer of proteins onto PVDF membranes (Millipore, IPVH00010), nonspecific binding was blocked by incubation with 5% skim milk, then membranes were incubated with primary antibodies against: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB12010454, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-caspase-8 (Cell Signaling Technologies [CST], 4927, 1:1000), anti-cleaved caspase-8 (CST, 8592, 1:1000), anticaspase-3 (CST, 9662, 1:1000), anti-cleaved caspase-3 (CST, 9661, 1:1000), anti-caspase-7 (CST, 9492, 1:1000), anti-cleaved caspase-7 (CST, 9491, 1:1000), anti-pRIPK3 (CST, 91702 S, 1:1000), anti-RIPK3 (ProSci, 2283, 1:1000), anti-pMLKL (CST, 37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-NLRP3 (AdipoGen, AG-20B-0044, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (1:1000) (ref. 88), anti-LDHA (Proteintech, 19987-1-AP, 1:1000), antiHMGB1 (Abcam, ab79823, 1:1000), anti-cleaved IL-1β (CST, 63124, 1:1000), anti-IL-1β (CST, 12426, 1:1000), anti-IL-18 (Abcam, ab207323, 1:1000), anti-ASC (Adipogen, AG-25B-0006, 1:1000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), and anti-β-actin (Proteintech, 66009-1-IG, 1:5000). .. Membranes were then washed and incubated with the appropriate horseradish peroxidase (HRP)–conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, anti-rabbit [111-035-047], 1:5000; anti-mouse [315-035-047], 1:5,000; and anti-rat [112-035-003], 1:5000) for 1 h. Proteins were visualized using Immobilon Forte Western HRP Substrate (Millipore, WBLUF0500), and membranes were developed with the GE Amersham Imager 600.
Western Blot:Article Title:
Article Snippet: .. ; Website: https://adipogen.com/ag-20b-0042-anti-caspase-1-p20-mouse-mab-casper-1.html 2. anti-caspase-11 (Novus Biologicals, NB120-10454): Species: Human, Mouse, Rat; Application: WB, ELISA, ICC, IHC, IP; Citation: Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation: G. Marine; Cell Rep. (2022). .. ; Website: https://www.novusbio.com/products/caspase-11-antibody-17d9_nb120-10454#reviews-publications 3. anti-caspase-3 (CST, #9662): Species: Human, Mouse, Rat; Application: WB, IHC, IP; Citation: Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death: L. Zhang; J. Exp.
Article Title:
Article Snippet: .. 3 nature portfolio | rep orting sum m ary April2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation Immunoblotting: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB120-10454, 1:1,000), anticaspase-3 (CST, #9662, 1:1000), anti-cleaved caspase-3 (CST, #9661, 1:1000), anti-caspase-7 (CST, #9492, 1:1000), anti-cleaved caspase-7 (CST, #9491, 1:1000), anti-caspase-8 (CST, #4927, 1:1000), anti-cleaved caspase-8 (CST, #8592, 1:1000), anti-pRIPK3 (CST, #91702S, 1:1000), anti-RIPK3 (ProSci, #2283, 1:1000), anti-pMLKL (CST, #37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-NLRP3 (Adipogen, AG-20B-0014-C100, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (Customized, Ref. 83, 1:1,000), anti-LDHA (Proteintech, 19987-1-AP, 1:1,000), anti-HMGB1 (Abcam, ab79823, 1:1,000), anti-cleaved IL-1 (CST, 63124, 1:1,000), anti-IL-1 (CST 12426, 1:1,000), anti-IL-18 (Abcam, ab207323, 1:1,000), anti-ASC (Adipogen, AG-25B-0006, 1:1,000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), anti-actin (Proteintech, 66009-1-IG, 1:5,000); anti-IgG (CST, #3900); secondary anti-rabbit (111-035-047), anti-mouse (315-035-047) or anti-rat (112-035-003) HRP antibodies from Jackson ImmunoResearch Laboratories. .. Microscopy: anti-ASC (Millipore, 04-147, 1:100), anti-RIPK3 (Millipore, MABC1595, 1:200), anti-CASP8 (CST, #4927, 1:200), antiNLRP3 (Adipogen, AG-20B-0014-C100, 1:200) or anti-NINJ1 (Customized, 1:200), Alexa Fluor 488 (ThermoFisher, A20181)-conjugated anti-ASC (Millipore, 04-147), Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (IgG) (Thermo Fisher Scientific, A11029, 1:200), Alexa Fluor 568-conjugated antibody against rat IgG (Thermo Fisher Scientific, A11077, 1:200), or Alexa Fluor 647- conjugated antibody against rabbit IgG (Thermo Fisher Scientific, A21245, 1:200) Antibodies were validated by their source company.
Enzyme-linked Immunosorbent Assay:Article Title:
Article Snippet: .. ; Website: https://adipogen.com/ag-20b-0042-anti-caspase-1-p20-mouse-mab-casper-1.html 2. anti-caspase-11 (Novus Biologicals, NB120-10454): Species: Human, Mouse, Rat; Application: WB, ELISA, ICC, IHC, IP; Citation: Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation: G. Marine; Cell Rep. (2022). .. ; Website: https://www.novusbio.com/products/caspase-11-antibody-17d9_nb120-10454#reviews-publications 3. anti-caspase-3 (CST, #9662): Species: Human, Mouse, Rat; Application: WB, IHC, IP; Citation: Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death: L. Zhang; J. Exp.
Immunocytochemistry:Article Title:
Article Snippet: .. ; Website: https://adipogen.com/ag-20b-0042-anti-caspase-1-p20-mouse-mab-casper-1.html 2. anti-caspase-11 (Novus Biologicals, NB120-10454): Species: Human, Mouse, Rat; Application: WB, ELISA, ICC, IHC, IP; Citation: Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation: G. Marine; Cell Rep. (2022). .. ; Website: https://www.novusbio.com/products/caspase-11-antibody-17d9_nb120-10454#reviews-publications 3. anti-caspase-3 (CST, #9662): Species: Human, Mouse, Rat; Application: WB, IHC, IP; Citation: Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death: L. Zhang; J. Exp.
Immunohistochemistry:Article Title:
Article Snippet: .. ; Website: https://adipogen.com/ag-20b-0042-anti-caspase-1-p20-mouse-mab-casper-1.html 2. anti-caspase-11 (Novus Biologicals, NB120-10454): Species: Human, Mouse, Rat; Application: WB, ELISA, ICC, IHC, IP; Citation: Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation: G. Marine; Cell Rep. (2022). .. ; Website: https://www.novusbio.com/products/caspase-11-antibody-17d9_nb120-10454#reviews-publications 3. anti-caspase-3 (CST, #9662): Species: Human, Mouse, Rat; Application: WB, IHC, IP; Citation: Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death: L. Zhang; J. Exp.
Membrane:Article Title:
Article Snippet: .. ; Website: https://adipogen.com/ag-20b-0042-anti-caspase-1-p20-mouse-mab-casper-1.html 2. anti-caspase-11 (Novus Biologicals, NB120-10454): Species: Human, Mouse, Rat; Application: WB, ELISA, ICC, IHC, IP; Citation: Endocytic membrane repair by ESCRT-III controls antigen export to the cytosol during antigen cross-presentation: G. Marine; Cell Rep. (2022). .. ; Website: https://www.novusbio.com/products/caspase-11-antibody-17d9_nb120-10454#reviews-publications 3. anti-caspase-3 (CST, #9662): Species: Human, Mouse, Rat; Application: WB, IHC, IP; Citation: Type I interferon signaling mediates Mycobacterium tuberculosis-induced macrophage death: L. Zhang; J. Exp.
Chromatin Immunoprecipitation:Article Title:
Article Snippet: .. 3 nature portfolio | rep orting sum m ary April2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation Immunoblotting: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB120-10454, 1:1,000), anticaspase-3 (CST, #9662, 1:1000), anti-cleaved caspase-3 (CST, #9661, 1:1000), anti-caspase-7 (CST, #9492, 1:1000), anti-cleaved caspase-7 (CST, #9491, 1:1000), anti-caspase-8 (CST, #4927, 1:1000), anti-cleaved caspase-8 (CST, #8592, 1:1000), anti-pRIPK3 (CST, #91702S, 1:1000), anti-RIPK3 (ProSci, #2283, 1:1000), anti-pMLKL (CST, #37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-NLRP3 (Adipogen, AG-20B-0014-C100, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (Customized, Ref. 83, 1:1,000), anti-LDHA (Proteintech, 19987-1-AP, 1:1,000), anti-HMGB1 (Abcam, ab79823, 1:1,000), anti-cleaved IL-1 (CST, 63124, 1:1,000), anti-IL-1 (CST 12426, 1:1,000), anti-IL-18 (Abcam, ab207323, 1:1,000), anti-ASC (Adipogen, AG-25B-0006, 1:1,000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), anti-actin (Proteintech, 66009-1-IG, 1:5,000); anti-IgG (CST, #3900); secondary anti-rabbit (111-035-047), anti-mouse (315-035-047) or anti-rat (112-035-003) HRP antibodies from Jackson ImmunoResearch Laboratories. .. Microscopy: anti-ASC (Millipore, 04-147, 1:100), anti-RIPK3 (Millipore, MABC1595, 1:200), anti-CASP8 (CST, #4927, 1:200), antiNLRP3 (Adipogen, AG-20B-0014-C100, 1:200) or anti-NINJ1 (Customized, 1:200), Alexa Fluor 488 (ThermoFisher, A20181)-conjugated anti-ASC (Millipore, 04-147), Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (IgG) (Thermo Fisher Scientific, A11029, 1:200), Alexa Fluor 568-conjugated antibody against rat IgG (Thermo Fisher Scientific, A11077, 1:200), or Alexa Fluor 647- conjugated antibody against rabbit IgG (Thermo Fisher Scientific, A21245, 1:200) Antibodies were validated by their source company.
Flow Cytometry:Article Title:
Article Snippet: .. 3 nature portfolio | rep orting sum m ary April2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation Immunoblotting: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB120-10454, 1:1,000), anticaspase-3 (CST, #9662, 1:1000), anti-cleaved caspase-3 (CST, #9661, 1:1000), anti-caspase-7 (CST, #9492, 1:1000), anti-cleaved caspase-7 (CST, #9491, 1:1000), anti-caspase-8 (CST, #4927, 1:1000), anti-cleaved caspase-8 (CST, #8592, 1:1000), anti-pRIPK3 (CST, #91702S, 1:1000), anti-RIPK3 (ProSci, #2283, 1:1000), anti-pMLKL (CST, #37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-NLRP3 (Adipogen, AG-20B-0014-C100, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (Customized, Ref. 83, 1:1,000), anti-LDHA (Proteintech, 19987-1-AP, 1:1,000), anti-HMGB1 (Abcam, ab79823, 1:1,000), anti-cleaved IL-1 (CST, 63124, 1:1,000), anti-IL-1 (CST 12426, 1:1,000), anti-IL-18 (Abcam, ab207323, 1:1,000), anti-ASC (Adipogen, AG-25B-0006, 1:1,000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), anti-actin (Proteintech, 66009-1-IG, 1:5,000); anti-IgG (CST, #3900); secondary anti-rabbit (111-035-047), anti-mouse (315-035-047) or anti-rat (112-035-003) HRP antibodies from Jackson ImmunoResearch Laboratories. .. Microscopy: anti-ASC (Millipore, 04-147, 1:100), anti-RIPK3 (Millipore, MABC1595, 1:200), anti-CASP8 (CST, #4927, 1:200), antiNLRP3 (Adipogen, AG-20B-0014-C100, 1:200) or anti-NINJ1 (Customized, 1:200), Alexa Fluor 488 (ThermoFisher, A20181)-conjugated anti-ASC (Millipore, 04-147), Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (IgG) (Thermo Fisher Scientific, A11029, 1:200), Alexa Fluor 568-conjugated antibody against rat IgG (Thermo Fisher Scientific, A11077, 1:200), or Alexa Fluor 647- conjugated antibody against rabbit IgG (Thermo Fisher Scientific, A21245, 1:200) Antibodies were validated by their source company.
Magnetic Resonance Imaging:Article Title:
Article Snippet: .. 3 nature portfolio | rep orting sum m ary April2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation Immunoblotting: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB120-10454, 1:1,000), anticaspase-3 (CST, #9662, 1:1000), anti-cleaved caspase-3 (CST, #9661, 1:1000), anti-caspase-7 (CST, #9492, 1:1000), anti-cleaved caspase-7 (CST, #9491, 1:1000), anti-caspase-8 (CST, #4927, 1:1000), anti-cleaved caspase-8 (CST, #8592, 1:1000), anti-pRIPK3 (CST, #91702S, 1:1000), anti-RIPK3 (ProSci, #2283, 1:1000), anti-pMLKL (CST, #37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-NLRP3 (Adipogen, AG-20B-0014-C100, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (Customized, Ref. 83, 1:1,000), anti-LDHA (Proteintech, 19987-1-AP, 1:1,000), anti-HMGB1 (Abcam, ab79823, 1:1,000), anti-cleaved IL-1 (CST, 63124, 1:1,000), anti-IL-1 (CST 12426, 1:1,000), anti-IL-18 (Abcam, ab207323, 1:1,000), anti-ASC (Adipogen, AG-25B-0006, 1:1,000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), anti-actin (Proteintech, 66009-1-IG, 1:5,000); anti-IgG (CST, #3900); secondary anti-rabbit (111-035-047), anti-mouse (315-035-047) or anti-rat (112-035-003) HRP antibodies from Jackson ImmunoResearch Laboratories. .. Microscopy: anti-ASC (Millipore, 04-147, 1:100), anti-RIPK3 (Millipore, MABC1595, 1:200), anti-CASP8 (CST, #4927, 1:200), antiNLRP3 (Adipogen, AG-20B-0014-C100, 1:200) or anti-NINJ1 (Customized, 1:200), Alexa Fluor 488 (ThermoFisher, A20181)-conjugated anti-ASC (Millipore, 04-147), Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (IgG) (Thermo Fisher Scientific, A11029, 1:200), Alexa Fluor 568-conjugated antibody against rat IgG (Thermo Fisher Scientific, A11077, 1:200), or Alexa Fluor 647- conjugated antibody against rabbit IgG (Thermo Fisher Scientific, A21245, 1:200) Antibodies were validated by their source company.
Biomarker Discovery:Article Title:
Article Snippet: .. 3 nature portfolio | rep orting sum m ary April2023 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used Validation Immunoblotting: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB120-10454, 1:1,000), anticaspase-3 (CST, #9662, 1:1000), anti-cleaved caspase-3 (CST, #9661, 1:1000), anti-caspase-7 (CST, #9492, 1:1000), anti-cleaved caspase-7 (CST, #9491, 1:1000), anti-caspase-8 (CST, #4927, 1:1000), anti-cleaved caspase-8 (CST, #8592, 1:1000), anti-pRIPK3 (CST, #91702S, 1:1000), anti-RIPK3 (ProSci, #2283, 1:1000), anti-pMLKL (CST, #37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-NLRP3 (Adipogen, AG-20B-0014-C100, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (Customized, Ref. 83, 1:1,000), anti-LDHA (Proteintech, 19987-1-AP, 1:1,000), anti-HMGB1 (Abcam, ab79823, 1:1,000), anti-cleaved IL-1 (CST, 63124, 1:1,000), anti-IL-1 (CST 12426, 1:1,000), anti-IL-18 (Abcam, ab207323, 1:1,000), anti-ASC (Adipogen, AG-25B-0006, 1:1,000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), anti-actin (Proteintech, 66009-1-IG, 1:5,000); anti-IgG (CST, #3900); secondary anti-rabbit (111-035-047), anti-mouse (315-035-047) or anti-rat (112-035-003) HRP antibodies from Jackson ImmunoResearch Laboratories. .. Microscopy: anti-ASC (Millipore, 04-147, 1:100), anti-RIPK3 (Millipore, MABC1595, 1:200), anti-CASP8 (CST, #4927, 1:200), antiNLRP3 (Adipogen, AG-20B-0014-C100, 1:200) or anti-NINJ1 (Customized, 1:200), Alexa Fluor 488 (ThermoFisher, A20181)-conjugated anti-ASC (Millipore, 04-147), Alexa Fluor 488-conjugated antibody against mouse immunoglobulin G (IgG) (Thermo Fisher Scientific, A11029, 1:200), Alexa Fluor 568-conjugated antibody against rat IgG (Thermo Fisher Scientific, A11077, 1:200), or Alexa Fluor 647- conjugated antibody against rabbit IgG (Thermo Fisher Scientific, A21245, 1:200) Antibodies were validated by their source company.
Binding Assay:Article Title: NINJ1 mediates inflammatory cell death, PANoptosis, and lethality during infection conditions and heat stress.
Article Snippet: .. Following electrophoretic transfer of proteins onto PVDF membranes (Millipore, IPVH00010), nonspecific binding was blocked by incubation with 5% skim milk, then membranes were incubated with primary antibodies against: anti-caspase-1 (AdipoGen, AG-20B-0044, 1:1000), anti-caspase-11 (Novus Biologicals, NB12010454, 1:1000), anti-GSDMD (Abcam, ab209845, 1:1000), anti-GSDME (Abcam, ab215191, 1:1000), anti-caspase-8 (Cell Signaling Technologies [CST], 4927, 1:1000), anti-cleaved caspase-8 (CST, 8592, 1:1000), anticaspase-3 (CST, 9662, 1:1000), anti-cleaved caspase-3 (CST, 9661, 1:1000), anti-caspase-7 (CST, 9492, 1:1000), anti-cleaved caspase-7 (CST, 9491, 1:1000), anti-pRIPK3 (CST, 91702 S, 1:1000), anti-RIPK3 (ProSci, 2283, 1:1000), anti-pMLKL (CST, 37333, 1:1000), anti-MLKL (Abgent, AP14272b, 1:1000), anti-NLRP3 (AdipoGen, AG-20B-0044, 1:1000), anti-ZBP1 (AdipoGen, AG-20B-0010, 1:1000), anti-NINJ1 (1:1000) (ref. 88), anti-LDHA (Proteintech, 19987-1-AP, 1:1000), antiHMGB1 (Abcam, ab79823, 1:1000), anti-cleaved IL-1β (CST, 63124, 1:1000), anti-IL-1β (CST, 12426, 1:1000), anti-IL-18 (Abcam, ab207323, 1:1000), anti-ASC (Adipogen, AG-25B-0006, 1:1000), anti-GAPDH (Santacruz, sc-166574 HRP, 1:10,000), and anti-β-actin (Proteintech, 66009-1-IG, 1:5000). .. Membranes were then washed and incubated with the appropriate horseradish peroxidase (HRP)–conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, anti-rabbit [111-035-047], 1:5000; anti-mouse [315-035-047], 1:5,000; and anti-rat [112-035-003], 1:5000) for 1 h. Proteins were visualized using Immobilon Forte Western HRP Substrate (Millipore, WBLUF0500), and membranes were developed with the GE Amersham Imager 600.
Cell-Signaling:Article Title: Deficiency of circadian clock gene Bmal1 exacerbates noncanonical inflammasome-mediated pyroptosis and lethality via Rev-erbα-C/EBPβ-SAA1 axis
Article Snippet: SAA1 ELISA kits were purchased from R&D Systems. .. The following antibodies were used in this study: anti-caspase-1 (AG-20B-0042, Adipogen), anti-NLRP3 (AG-20B-00140, Adipogen), anti-mouse IL-1β (AF-401-NA, R&D Systems), anti-caspase-11 (NB120-10454, Novus Biologicals), anti-Bmal1 (93806, Abcam), anti-GSDMD (209845, Abcam), anti-cathepsin B (AF965, R&D Systems), anti-cleaved caspase-3 (9579, Cell Signaling Technology), anti-fibrin (MVI-1007, MERU-VasImmune), anti-phospho-STAT1 (#7649, Cell Signaling Technology), anti-STAT1 (#14994, Cell Signaling Technology), anti-β-actin (sc-47778, Santa Cruz Biotechnology), anti-LAMP-1 (sc-19992, Santa Cruz Biotechnology) and anti-C/EBPβ (sc-7962, Santa Cruz Biotechnology). ..
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