Cells were harvested and lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing protease inhibitor cocktail (Sigma-Aldrich). Proteins samples were separated using 10–15% SDS-PAGE and then transferred to membranes (Millipore, Burlington, MA, USA), which were then probed with primary and secondary antibodies. Membranes were developed using a chemiluminescence solution (GE Healthcare, Chicago, IL, USA) in a LAS-4000 Lumino-imaging unit (Fujifilm, Tokyo, Japan). Immunoblot band intensities were quantified using NIH ImageJ software (Version 1.53t, Fujifilm), and results were presented as intensity ratios versus β-actin. The antibodies used were as follows; anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti-human iNOS (Santa Cruz Biotechnology), anti-human TLR2 (Santa Cruz Biotechnology), anti-human TLR4 (Santa Cruz Biotechnology), NF-κB (Cell Signaling Technology), MAPK Family antibody sampler kit (Cell Signaling Technology), anti-human AIM2 (Cell Signaling Technology), anti-human ASC (Cell Signaling Technology), pro-Caspase-1 (Cell Signaling Technology), anti-human IL-1β (R&D Systems, Minneapolis, MN, USA), Beclin1 (Santa Cruz Biotechnology), ATG5 (Santa Cruz Biotechnology), and LC3 (Cell Signaling Technology).
Anti human il 1β
Manufactured by R&D Systems
Sourced in United States
Anti-human IL-1β is a recombinant protein that binds and neutralizes human interleukin-1 beta (IL-1β). IL-1β is a pro-inflammatory cytokine that plays a crucial role in immune and inflammatory responses.
Automatically generated - may contain errors
Lab products found in correlation
Anti β actin, by Santa Cruz Biotechnology (3 mentions)
Anti human aim2, by Cell Signaling Technology (3 mentions)
Anti human asc, by Cell Signaling Technology (3 mentions)
Anti mouse il 1β, by R&D Systems (3 mentions)
Chemiluminescence solution, by GE Healthcare (2 mentions)
Las 4000 lumino imaging unit, by Fujifilm (2 mentions)
Pro caspase 1, by Cell Signaling Technology (2 mentions)
Ripa buffer, by Cell Signaling Technology (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
β actin, by Fujifilm (2 mentions)
Anti mouse caspase 1, by Abcam (2 mentions)
Anti mouse nlrp3, by Cell Signaling Technology (2 mentions)
Anti human caspase 1, by Abcam (2 mentions)
Anti human nlrp3, by Abcam (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Mapk family antibody sampler kit, by Cell Signaling Technology (1 mentions)
Beclin 1, by Santa Cruz Biotechnology (1 mentions)
Nf κb, by Cell Signaling Technology (1 mentions)
Z vad fmk, by R&D Systems (1 mentions)
Z wehd fmk, by R&D Systems (1 mentions)
8 protocols using anti human il 1β
1
Immunoblot Analysis of Inflammatory Pathways
2
Inflammasome Activation Pathway Protocols
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Phorbol 12-mystristate 13-acetate (PMA), oxidised ATP (oxATP), KCl, and trichloroacetate (TCA) were purchased from Sigma (St. Louis, MO, USA). CA-074 methyl ester (CA-074Me) was obtained from Calbiochem (San Diego, CA, USA). Z-VAD-FMK(pan-caspase inhibitor), Z-WEHD-FMK(caspase-1 inhibitor), anti-human IL-1β, and anti-human NLRP3 were supplied by R&D Systems (Minneapolis, MN). Anti-human ASC, anti-human AIM2, and anti-human caspase-1 antibodies were acquired from Cell Signaling Technology (Beverly, MA, USA). The anti-human NLRC4 and anti-β-actin antibodies were procured from Santa Cruz (Santa Cruz, CA, USA).
3
Inflammatory Pathway Activation in Viral Lung Infection
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Lungs were harvested one day post-infection, dissolved in lysis buffer and homogenized for Western blot assay using anti-mouse IL-1β (R&D Systems), anti-mouse caspase-1 (Abcam, Cambridge, MA), anti-mouse NLRP3 (Cell Signaling Technology), and anti-β actin (Millipore Sigma, Burlington, MA). RV-A1B- and RV-C15-infected THP-1-derived macrophages were harvested one-day post-infection, dissolved in lysis buffer for Western blot assay using anti-human IL-1β (R&D Systems), anti-human caspase-1 (Abcam), anti-human NLRP3 (Abcam).
4
Inflammatory Pathway Activation in Viral Lung Infection
5
Western Blot Analysis of Inflammatory Markers
6
Western Blot Analysis of Inflammasome Proteins
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Cells were harvested and lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) containing protease inhibitor cocktail (Sigma-Aldrich). Protein samples were separated using 10–15% SDS-PAGE and then transferred to membranes (MilliporeSigma, Burlington, MA, USA), which were then probed with primary and secondary antibodies. Membranes were developed using a chemiluminescence solution (GE Healthcare, Chicago, IL, USA) in a LAS-4000 Lumino-imaging unit (Fujifilm, Tokyo, Japan). Immunoblot band intensities were quantified using NIH ImageJ 1.54f software (Fujifilm), and the results are presented as intensity ratios versus β-actin. The antibodies used were as follows: anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA), anti-human AIM2 (Cell Signaling Technology), anti-human ASC (Cell Signaling Technology), pro-Caspase-1 (Cell Signaling Technology), anti-human IL-1β (R&D Systems, Minneapolis, MN, USA), and NLRP3 (NOVUS biologicals, Centennial, CO, USA)
7
Investigating Cytokine Responses in LPS-Stimulated Cells
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LPS (Escherichia coli 026:B6), adenosine 5′-triphosphate disodium salt hydrate (A2383; ATP), nigericin (N7143), and penicillin–streptomycin (Pen/Strep, P4333) were purchased from Sigma-Aldrich. Human IFN Beta (Beta 1a, Mammalian Expressed) (11415-1) and Human IFN Alpha Hybrid (Universal Type I IFN) (11200-1) were obtained from PBL Assay Science. Phosphatase inhibitor cocktail (78420) was sourced from Thermo Fisher Scientific, and FBS was obtained from Gibco.
Primary antibodies for Western blot analysis and their final concentrations were as follows: anti-human IL-1β (0.1 μg/ml, goat polyclonal, AF-201-NA; R&D Systems) and anti-β-actin HRP (0.2 μg/ml, mouse monoclonal, A3854; Sigma-Aldrich).
Primary antibodies for Western blot analysis and their final concentrations were as follows: anti-human IL-1β (0.1 μg/ml, goat polyclonal, AF-201-NA; R&D Systems) and anti-β-actin HRP (0.2 μg/ml, mouse monoclonal, A3854; Sigma-Aldrich).
8
Mast Cell Activation by Tumor Secretomes
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Mast cells were seeded at 5 × 105 cells per ml with 50% (v/v) medium 2 and cell culture supernatants. NHM, A375, and RPMI-7951 cell culture supernatants were used as well as medium alone [50% (v/v) medium 2 and supplemented DMEM] for control. In some cases, MCs were incubated for 30 min with neutralizing antibodies or appropriate isotype control antibodies before treatments. The following blocking antibodies were used with their respective isotype controls: anti-human IL-33 (50 μg/ml) (AF3625), goat IgG control (AB-108-C), anti-human IL-1β (50 μg/ml) (clone: 8516, MAB201), and mouse IgG1 isotype control (MAB002R) antibodies all from R&D Systems, Mineapolis, USA and anti-human TGF-β1 (100 μg/ml) (clone: 1D11.16.8) and mouse IgG1 isotype control (clone: MOPC-21) from Bio X Cell, USA.
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