All chemicals used were obtained from commercial sources and were of analytical grade. Media components, fine chemicals and reagents were purchased from Sigma Aldrich (St. Louis, USA), HiMedia (Mumbai, India), Merck Millipore India Ltd (Mumbai, India), USB Corporation (Ohio, USA) or Difco, USA. Cysteine and cystine were also purchased from Sigma Aldrich (St. Louis, USA). HA-Tag (6E2) Mouse monoclonal antibody and horse anti-mouse HRP-linked antibody were bought from Cell Signaling Technology (Danvers, MA, USA). Alexa Flour® 488 conjugated goat anti-mouse antibody was obtained from Molecular probes (Eugene, Oregon, USA). Cysteine stock solutions that were prepared fresh were made by dissolving the required amount of Cysteine in 1 ml of de-ionized water, which was then filter-sterilized using 0.2 μ filter membrane. cystine stock solutions were prepared by dissolving the required amount of cystine in 1 ml of deionized water along with 25 µL of concentrated HCl, filter-sterilized using 0.2 μ filter membrane. 14C cystine and custom synthesized 35S cystine was procured from Perkin Elmer Ltd. (Boston, USA).
Horse anti mouse hrp linked antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Horse anti-mouse HRP-linked antibody is a secondary antibody that binds to mouse primary antibodies, enabling detection and visualization in immunoassay applications. The antibody is conjugated with horseradish peroxidase (HRP), which can catalyze a colorimetric or chemiluminescent reaction for signal amplification.
Automatically generated - may contain errors
Lab products found in correlation
Ha tag 6e2 mouse monoclonal antibody, by Cell Signaling Technology (2 mentions)
Alexa flour 488 conjugated goat anti mouse antibody, by Thermo Fisher Scientific (2 mentions)
Hrp linked goat anti rabbit antibody, by Cell Signaling Technology (2 mentions)
Lipopolysaccharides (lps), by Merck Group (2 mentions)
Anti flag, by Merck Group (2 mentions)
Anti actin conjugated to horseradish peroxidase, by Merck Group (2 mentions)
Anti caspase 1, by Cell Signaling Technology (2 mentions)
Anti il 1β, by Cell Signaling Technology (2 mentions)
Gel extraction kit, by Qiagen (2 mentions)
Restriction enzyme, by New England Biolabs (2 mentions)
Plasmid miniprep columns, by Qiagen (2 mentions)
Vent dna polymerase, by New England Biolabs (2 mentions)
Cysteine, by Merck Group (1 mentions)
Cystine, by Merck Group (1 mentions)
14c cystine, by PerkinElmer (1 mentions)
35s cystine, by PerkinElmer (1 mentions)
Sytox green nucleic acid stain, by Thermo Fisher Scientific (1 mentions)
Nigericin, by Merck Group (1 mentions)
Anti bcl 2, by Santa Cruz Biotechnology (1 mentions)
Anti myc tag, by Cell Signaling Technology (1 mentions)
5 protocols using horse anti mouse hrp linked antibody
1
Cysteine and Cystine Preparation Protocol
2
Immunoblotting Analysis of Pyroptosis Proteins
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Antibodies used for immunoblotting analysis were the following: anti-IL-1β (#12703), anti-caspase-1 (#3866), anti-caspase-4 (#4450), anti-caspase-5 (#46680), anti-MLKL (#14993), anti-phospho-MLKL (#91689), anti-RIP-3 (#13526), anti-phospho-RIP3 (#93654) and anti-Myc-tag (#2278) (Cell Signaling Technology); and anti-Flag (Sigma-Aldrich, F1804), anti-Bcl-2 (Santa Cruz Biotechnology, sc-783), anti-actin conjugated to horseradish peroxidase (Sigma-Aldrich, A3854), anti-GAPDH conjugated to horseradish peroxidase (Proteintech, HRP-60004), goat anti-rabbit HRP-linked antibody (#7074), and horse anti-mouse HRP-linked antibody (#7076) (Cell Signaling Technology). Anti-GSDMD was from (Novusbio, NBP2-33422). The LPS, nigericin and Z-VAD-FMK caspase inhibitor were purchased from Sigma-Aldrich. SYTOX Green nucleic acid stain was purchased from Invitrogen. Smac mimetic was from Tocris. Caspase-1, caspase-3 and Bcl-2 recombinant proteins were from EMD Millipore. Human TNFα was from R&D System. Smac mimic was from APExBIO.
3
Molecular Cloning and Protein Purification
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All chemicals used in the present study were of either analytical or molecular biology grades and were obtained from commercial sources. Media components were purchased from Difco (USA). Oligonucleotides were purchased from Sigma (India) and IDT (USA). Restriction enzymes, Vent DNA polymerase, and other DNA-modifying enzymes were obtained from New England Biolabs (USA). Gel-extraction kits, plasmid miniprep columns, and the Ni-NTA agarose resin were obtained from Qiagen (Germany). Hybridization nitrocellulose membrane (filter type 0.45 μm) and Luminata forte Western horseradish peroxidase (HRP) substrate was obtained from Millipore (India). Anti-His mouse monoclonal antibody (27E8) and horse anti-mouse HRP-linked antibody were procured from Cell Signaling Technology (USA). Anti-GSH mouse monoclonal antibody (ab19534) was from Abcam (UK). Alexa Fluor 488–conjugated goat anti-mouse antibody was obtained from Molecular Probes (USA). Coelentrazine was purchased from Promega (USA).
4
Immunoblot Analysis of Inflammasome Components
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Antibodies used for immunoblot analysis were the following: anti-NLRP3, anti-IL-1β, anti-caspase-1, anti-caspase-9 and anti-cytochrome c (Cell Signaling); and anti-Myc (Clontech Laboratories), anti-Flag (Sigma-Aldrich), anti-HA (Cell Signaling), anti-ASC (Santa Cruz Biotechnology), anti-actin conjugated to horseradish peroxidase (Sigma-Aldrich), goat anti-rabbit HRP-linked antibody, and horse anti-mouse HRP-linked antibody (Cell Signaling). Purified cytochrome c protein (Sigma-Aldrich), Profect-P1-lipid based protein delivery reagent (Targeting Systems), cardiolipin beads (Echelon), and the mitochondrial fractionation kit (Active Motif) were purchased and used as directed by the manufactures. The LPS, ATP, and Poly (dA-dT) were purchased from Sigma-Aldrich.
5
Detailed Biochemical Experimental Protocols
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All the chemicals used in this study were analytical grade and obtained from commercial sources. Media components were purchased from Difco (Detroit, MI) Sigma Aldrich, (St. Louis, MO), HiMedia, (Mumbai, India), Merck India Ltd (Mumbai, India), and USB Corporation (Cleveland, OH). Oligonucleotides were purchased from Sigma India. Restriction enzymes, Vent DNA polymerase, and other DNA-modifying enzymes were obtained from New England Biolabs (Beverly, MA). DNA sequencing kit (ABI PRISM 310 XL with dye termination cycle sequencing ready reaction kit) was obtained from Perkin Elmer, (Norwalk, CT). Gel-extraction kits and plasmid miniprep columns were obtained from QIAGEN (Valencia, CA) or Sigma (St. Louis, MO). [35S] GSH (specific activity 1000 Ci mmol-1) was purchased from Bhabha Atomic Research Centre, Mumbai, India. HA-Tag (6E2) mouse monoclonal antibody and horse anti-mouse HRP-linked antibody were bought from Cell Signaling (Danvers, MA). Alexa Flour 488 conjugated goat anti-mouse antibody was obtained from Molecular Probes (Eugene, OR). Hybond ECL (nitrocellulose) membrane and ECL plus Western blotting detection reagents were purchased from Amersham Biosciences (UK).
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