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Human il 1β

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Human IL-1β is a recombinant human cytokine that functions as a pro-inflammatory mediator. It is a member of the interleukin-1 family and is primarily produced by activated macrophages, dendritic cells, and other immune cells in response to various stimuli. This protein plays a crucial role in the regulation of immune and inflammatory responses.

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Lab products found in correlation

Human il 23, by R&D Systems (2 mentions) Mouse il 1β, by R&D Systems (2 mentions) Il 1β, by R&D Systems (2 mentions) Interleukin 6 (il 6), by R&D Systems (2 mentions) Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions) Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Nigericin, by Merck Group (1 mentions) Lps from escherichia coli serotype 055 b5, by Merck Group (1 mentions) Human pro il 1β, by Sino Biological (1 mentions) Bapta am, by Thermo Fisher Scientific (1 mentions) Goat anti human il 1β antibody, by R&D Systems (1 mentions) Rabbit anti asc antibody, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488 conjugated goat anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions) Human il 8, by BD (1 mentions) Nigericin, by Cayman Chemical (1 mentions) Vildagliptin, by Cayman Chemical (1 mentions) Saxagliptin, by LGC (1 mentions) Sitagliptin, by Merck Group (1 mentions) Lipopolysaccharides (lps), by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Human TNF-α and IL-1β ELISA kit Recombinant human and mouse IL-1β Human IL-1β/IL-1F2 DuoSet ELISA Development Kit Human IL-1β or TNF-α Recombinant human IL-1β (201-LB) Recombinant human interleukin (IL)-1β Human IL-1β/IL-1F2 DuoSet ELISA kit Human IL-1β/IL-1F2 DuoSet ELISA Recombinant human IL-1α or IL-1β Human IL-1β/IL-1F2 DuoSet

28 protocols using human il 1β

1

Inflammasome Activation by LPS and Nigericin

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LPS from Escherichia coli serotype 055:B5 (Toll like receptors 2/4) and nigericin were purchased from Sigma. The recombinant proteins used were human pro-IL-1β, human calmodulin (both from Sino Biological, Philadelphia, PA), and human IL-1β (R&D Systems, Minneapolis, MN). The calcium chelator BAPTA-AM was purchased from Life Technologies, and the calmodulin inhibitors E6 berbamine and W7 were purchased from Enzo Life Sciences (Exeter, UK) and Santa Cruz Biotechnology, respectively. For Western blot analysis, the primary antibodies used were a goat anti-human IL-1β antibody (R&D Systems) or a rabbit anti-human caspase-1 (p10) antibody (Santa Cruz Biotechnology). The secondary antibodies used were a sheep anti-mouse IgG antibody (AbD Serotech, Kidlington, UK) or a goat anti-rabbit antibody (Dako, Copenhagen, Denmark). For immunofluorescence analysis, the primary antibodies used were a rabbit anti-ASC antibody (Santa Cruz Biotechnology), a rabbit anti-calmodulin antibody (Abcam, Cambridge, UK), or a goat anti-human IL-1β antibody (R&D Systems). The secondary antibodies used were an Alexa Fluor 488-conjugated goat anti-rabbit IgG antibody or an Alexa Fluor 594-conjugated rabbit anti-goat IgG antibody (both from Life Technologies).
Ainscough J.S., Gerberick G.F., Kimber I, & Dearman R.J. (2015). Interleukin-1β Processing Is Dependent on a Calcium-mediated Interaction with Calmodulin. The Journal of Biological Chemistry, 290(52), 31151-31161.
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2

Cytokine and hs-CRP Quantification in Patient Serum

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For analyses of IL-1 β, IL-6, IL-8, and TNF-α, the blood samples were collected from the patients at 8 am after an overnight fast. The serum samples were immediately centrifuged and were stored at -80°C until further determination. IL-1β, IL-6, and IL-8 were determined with the commercial enzyme-linked immunosorbent assay kits (human IL-1β: R&D Systems INC., Minnesota, USA; human IL-6: high sensitivity enzyme-linked immunosorbent assay, Immunotech S. A, Marseille, France; human IL-8: enzyme-linked immunosorbent assay; BD Bioscience, La Jolla, CA) as previously described [17 (link), 18 (link)]. The serum samples were stored in -80°C until further determination, and the serum levels of TNF-α were determined by a high sensitivity enzyme-linked immunosorbent assay kit (Biosource, International, Inc.) according to the manufacturer's instructions [19 (link)]. The serum levels of high sensitive C reaction protein (hs-CRP) were analyzed with a commercially available assay kit (hs-CRP test kits, Millipore Corp, Billerica, MA, USA), and the levels of hs-CRP were expressed as mg/L.
Li X., Guo D., Chen Y., Hu Y, & Zhang F. (2020). Effects of Altered Levels of Pro- and Anti-Inflammatory Mediators on Locations of In-Stent Reocclusions in Elderly Patients. Mediators of Inflammation, 2020, 1719279.
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3

Caspase Activation and Pyroptosis Assays

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LPS was purchased from Santa Cruz Biotechnology, nigericin, and vildagliptin and Ac-YVAD-CMK from the Cayman Chemical Company, PMA and sitagliptin from Sigma, Ala-Pro-AFC from Bachem, saxagliptin from Toronto Research Chemicals, and Z-VAD-FMK and etoposide from Enzo Life Sciences. Val-boroPro45 (link), 1G24424 (link), FP-biotin15 (link), L-allo-Ile-isoindoline14 (link), and L-allo-Ile-thiazolidine14 (link) were synthesized according to previously published protocols. For cell culture experiments, Val-boroPro was resuspended in DMSO containing 0.1% TFA to prevent compound cyclization. Antibodies used include: human caspase-1 (#2225, Cell Signaling Technology), mouse caspase-1 (clone Casper-1, Adipogen), caspase-3 (clone 8G10, Cell Signaling Technology), human caspase-4 (clone 4B9, Santa Cruz), human caspase-5 (clone D3G4W, Cell Signaling Technology), caspase-7 (clone D2Q3L, Cell Signaling Technology), human IL-1β (Clone 2805, R&D Systems), mouse IL-1β (clone D4T2D, Cell Signaling Technology), IL-1α (#AF-200, R&D Systems), IL-18 (#AF2548, R&D Systems), GAPDH (clone 14C10, Cell Signaling Technology), DPP7 (Clone 398024, R&D Systems), DPP8 (ab42076, Abcam), DPP9 (ab42080, Abcam), PARP (#9542, Cell Signaling Technology), GSDMD (NBP2-33422, Novus Biologicals), DPP4 (#11D7, GeneTex), FAP (ABT11, Millipore), and SCPEP1 (SAB2700267, Sigma).
Okondo M.C., Johnson D.C., Sridharan R., Go E.B., Chui A.J., Wang M.S., Poplawski S.E., Wu W., Liu Y., Lai J.H., Sanford D.G., Arciprete M.O., Golub T.R., Bachovchin W.W, & Bachovchin D.A. (2016). DPP8/9 inhibition induces pro-caspase-1-dependent monocyte and macrophage pyroptosis. Nature chemical biology, 13(1), 46-53.
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4

Investigating Cytokine Regulation by KW3110

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Cells were seeded and incubated overnight in 24-well plates (J774A.1 cells, 1 × 105 cells/well; human monocytes, 2.5 × 105 cells/well), and treated with KW3110 (1.25–5 μg/mL for J774A.1 cells and 100 μg/mL for human monocytes) for 24 h at 37 ˚C. To measure IL-1β, the cells were treated with 1 μg/mL of IL-10 or primed with 10 μg/mL of LPS for 4 h, and stimulated with 2 mM ATP for 1 h. For the supernatant transferal experiments, in Fig 1B, the supernatants collected from KW3110-treated J774A.1 cells were transferred to other cells which were then primed with 10 μg/mL of LPS for 4 h and stimulated with 2 mM ATP for 1 h. For the phagocytosis blocking experiments, 1 μg/mL of cytochalasin D was added 30 min prior to KW3110 treatment of J774A.1 cells. The supernatants were then collected and centrifuged at 5,000 rpm for 2 min. Cytokine levels were measured with commercial enzyme-linked immunosorbent assay (ELISA) kits (mouse and human IL-10, BD Biosciences, San Jose, CA, USA; mouse IL-1β, eBioscience, San Diego, CA, USA; human IL-1β, R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s instructions.
Yamazaki T., Ohshio K., Sugamata M, & Morita Y. (2020). Lactic acid bacterium, Lactobacillus paracasei KW3110, suppresses inflammatory stress-induced caspase-1 activation by promoting interleukin-10 production in mouse and human immune cells. PLoS ONE, 15(8), e0237754.
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5

Cytokine-induced Inflammation in ARPE-19 Cells

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ARPE-19 human RPE cells obtained from ATCC (Manassas, VA) were grown to confluence as we reported earlier [8 (link), 9 (link)]. The cells were treated with the proinflammatory cytokines IFN-γ (10 u/ml), IL-1β (1 ng/ml) and TNF-α (1 ng/ml) in the absence of serum for 16 h unless otherwise indicated. The cells were pre-incubated with Resveratrol (50 μM) for 4 hours also in the absence of serum prior to the cytokine treatment when required. Resveratrol was obtained from Sigma-Aldrich, St. Louis, MO and dissolved in ethanol before adding to cell culture medium. Equal volume of ethanol was added to controls. Human IL-1β was purchased from R&D Systems, Minneapolis, MN while TNF-α and IFN-γ were from Roche Applied Science, Indianapolis, IN.
Kutty R.K., Samuel W., Abay R., Cherukuri A., Nagineni C.N., Duncan T., Jaworski C., Vijayasarathy C, & Redmond T.M. (2015). Resveratrol Attenuates CXCL11 Expression Induced by Proinflammatory Cytokines in Retinal Pigment Epithelial Cells. Cytokine, 74(2), 335-338.
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6

Immune Response Modulation by TLR Agonists

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Cells were stimulated as indicated with TLR7-stimulatory RNA (5’-ACUG1CG1AG1CUU-X-UUCG1AG1CG1UCA-5’, G1 is 7-deazaguanosine, X is 1,2,3-propanetriol) from Idera Pharmaceuticals, R848 (InvivoGen), TLR13-stimulatory RNA (ORN Sa19, InvivoGen), LPS (InvivoGen), CpG1826 PTO (Metabion), CpG2006 (Metabion), heat-killed GBS (kind gift from Dr. G. Teti), human TNF (R&D Systems) or human IL-1β (R&D Systems). Transient transfections were performed using Lipofectamine 2000 transfection reagent (Thermo Fisher Scientific). Where indicated, cells were pre-incubated with Bafilomycin A1 (biomol). Cytokines were measured using commercially available ELISA kits (R&D Systems and BD Biosciences).
Pelka K., Bertheloot D., Reimer E., Phulphagar K., Schmidt S.V., Christ A., Stahl R., Watson N., Miyake K., Hacohen N., Haas A., Brinkmann M.M., Marshak-Rothstein A., Meissner F, & Latz E. (2018). The chaperone UNC93B1 regulates toll-like receptor stability independent of endosomal TLR transport. Immunity, 48(5), 911-922.e7.
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7

Caspase-1 and Caspase-4 Mediated IL-1β Cleavage

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5 U of full-length human caspase-1 (Abcam) or caspase-4 (Abcam) recombinant proteins were incubated with 2 μg of recombinant human pro-IL-1β (Sino Biological Inc.) in caspase activity buffer (200 mM NaCl, 50 mM HEPES, pH 8.0, 50 mM KCl). The protein mix was incubated at 37°C for 0, 6, and 24 h. Cleavage of recombinant proteins was analysed by Western blot using standard methods (Groß, 2012 ) and the following reagents: antibodies against the caspase-4 large subunit (4B9, mouse monoclonal antibody, 1:1,000; Santa Cruz Biotechnology), human IL-1β (goat polyclonal antibody, 1:1,000; R&D Systems), caspase-1 (D7F10, rabbit monoclonal, 1:1,000; Cell Signalling Technology).
Chan A.H., Burgener S.S., Vezyrgiannis K., Wang X., Acklam J., Von Pein J.B., Pizzuto M., Labzin L.I., Boucher D, & Schroder K. (2023). Caspase-4 dimerisation and D289 auto-processing elicit an interleukin-1β-converting enzyme. Life Science Alliance, 6(10), e202301908.
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8

Cytokine Profiling of Microglial Cells

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Cell culture supernatants collected from microglial cells at 24h and 48h post ssRNA40, ssRNA41 or vehicle treatment were used for quantification of cytokines using ELISA. Human IL-1β (R&D systems Cat# DLB50), human IL-18 (eBioscience Cat# BMS267–2), humanIL-1 alpha (eBioscience Cat# BMS243–2), human TNF-alpha (eBioscience Cat# BMS223–4), human complement C1q (Abcam Cat# ab170246) production were quantified by ELISA in these culture supernatants. These cell culture supernatants were also analyzed for relative levels of selected cytokines and chemokines using a membrane-based antibody array (R&D Systems Cat# ARY005B) following manufacturer’s instructions.
Rawat P., Diedrich C.T, & Spector S.A. (2018). Human Immunodeficiency Virus Type-1 single-stranded RNA activates the NLRP3 inflammasome and impairs autophagic clearance of damaged mitochondria in human microglia. Glia, 67(5), 802-824.
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9

Stimulation of NKp44+ ILCs from Rhesus Macaques

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NKp44+ ILCs from MLN of SIV-naïve rhesus macaques were stimulated overnight with either rhesus macaque IL-12 (50 ng/ml), human IL-1β (50 ng/ml), human IL-2 (1000 IU/ml) human IL-15 (50 ng/ml), or human IL-23 (50 ng/ml) (all from R&D Systems). After culture, cells were analyzed for intracellular expression of caspase-3 or RORγt (clone AFKJS-9, eBioscience).
Li H., Richert-Spuhler L.E., Evans T.I., Gillis J., Connole M., Estes J.D., Keele B.F., Klatt N.R, & Reeves R.K. (2014). Hypercytotoxicity and Rapid Loss of NKp44+ Innate Lymphoid Cells during Acute SIV Infection. PLoS Pathogens, 10(12), e1004551.
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10

Cytokine and Terminal Complement Assay

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Cytokine release was measured in supernatants using the Human IL-1β or IL-18 DuoSet ELISA kit (R&D) following manufacturer’s instructions.
The amount of MAC, also known as terminal complement complex (TCC), was measured in cell lysates using a human TCC ELISA kit (HycultBiotech) following manufacturer’s instructions.
Diaz-del-Olmo I., Worboys J., Martin-Sanchez F., Gritsenko A., Ambrose A.R., Tannahill G.M., Nichols E.M., Lopez-Castejon G, & Davis D.M. (2021). Internalization of the Membrane Attack Complex Triggers NLRP3 Inflammasome Activation and IL-1β Secretion in Human Macrophages. Frontiers in Immunology, 12, 720655.
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