Il 1β

Manufactured by Bio-Rad

Sourced in United States

IL-1β is a recombinant human interleukin-1 beta protein. Interleukin-1 beta is a pro-inflammatory cytokine that plays a central role in the regulation of immune and inflammatory responses.

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Lab products found in correlation

Tnf α, by Bio-Rad (7 mentions) Il 10, by Bio-Rad (5 mentions) Interleukin 6 (il 6), by Bio-Rad (5 mentions) Ifn γ, by Bio-Rad (5 mentions) G csf, by Bio-Rad (2 mentions) Il 17a, by Bio-Rad (2 mentions) Interleukin 4 (il 4), by Bio-Rad (2 mentions) Il 17, by Bio-Rad (2 mentions) Lps from e coli 0111 b4, by InvivoGen (1 mentions) Lsm lymphocyte separation medium, by MP Biomedicals (1 mentions) Il 1β, by R&D Systems (1 mentions) Tnf α, by R&D Systems (1 mentions) Interleukin 6 (il 6), by R&D Systems (1 mentions) Il 10, by R&D Systems (1 mentions) Creatinine, by Thermo Fisher Scientific (1 mentions) Troponin t, by R&D Systems (1 mentions) Manager 6, by Bio-Rad (1 mentions) Gm csf, by Bio-Rad (1 mentions) Eotaxin, by Bio-Rad (1 mentions) Ip 10, by Bio-Rad (1 mentions)

13 protocols using il 1β

Cytokine Profiling of LPS-Stimulated PBMCs

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On day 8 of the Control/baseline diet and on the 22^nd day of each experimental diet (HPA and HOA), blood was collected in the fasted state for measurement of cytokines in plasma and from LPS-stimulated, PBMCs. We enriched lymphocytes and monocytes from blood (PBMCs) using LSM Lymphocyte Separation Medium (MP Biomedicals, Solon, OH), and the mononuclear cell layer was collected, washed, and counted. Freshly-collected cells were plated at 10⁶ cells/ml and treated in duplicate under control conditions or were stimulated by the addition of 1ng/ml ultra-pure lipopolysaccharide (LPS; from E. coli 0111:B4, InvivoGen, San Diego, CA). After 24-hr., cell-free supernatants were collected and stored at −80° C until analysis. After pilot studies were conducted demonstrating which cytokines were responsive to LPS stimulation in our assays, the following cytokines were measured in these supernatants using a Luminex-based assay: TNFα, IL-1β, IL-6, and IL-10 (Bio-Rad, Hercules, CA) [8 (link)]. IL-18 was measured by ELISA (R&D Systems, Minneapolis, MN).
The following cytokines were measured in plasma using a Luminex-based assay: IL-1β, IL-6, IL-8, IL-10, and TNFα (R&D Systems, Minneapolis, MN).

Kien C.L., Bunn J.Y., Fukagawa N.K., Anathy V., Matthews D.E., Crain K.I., Ebenstein D.B., Tarleton E.K., Pratley R.E, & Poynter M.E. (2015). Lipidomic evidence that lowering the typical dietary palmitate to oleate ratio in humans decreases the leukocyte production of pro-inflammatory cytokines and muscle expression of redox-sensitive genes *, †, ‡. The Journal of nutritional biochemistry, 26(12), 1599-1606.

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Biomarker Analysis in Rat Serum

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A total of 2 ml peripheral venous blood was obtained from experimental rats after 4 weeks of treatment. Serum was obtained following centrifugation (6,000 × g; 15 min at 4°C). C reactive protein (CRP; cat. no. MCRP00; Bio-Rad Laboratories, Inc.), TNF-α (cat. no. RTA00; Bio-Rad Laboratories, Inc.), IL-6 (cat. no. R6000B; Bio-Rad Laboratories, Inc.), IL-1β (cat. no. DY501; Bio-Rad Laboratories, Inc.), brain natriuretic peptide (BNP; cat. no. MA134237; Thermo Fisher Scientific, Inc.), creatinine (cat. no. KGE005; R&D Systems, Inc.) and troponin T (cat. no. MAB1874; R&D Systems, Inc.) levels were analyzed using ELISA kits according to the manufacturers' instructions.

He L., Wang T., Chen B.W., Lu F.M, & Xu J. (2019). Puerarin inhibits apoptosis and inflammation in myocardial cells via PPARα expression in rats with chronic heart failure. Experimental and Therapeutic Medicine, 18(5), 3347-3356.

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Serum Cytokine Detection Protocol

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For serum cytokine detection, blood samples were collected together with CRP and PCT samples at time points 0, 12, 24, 48, 72, and 96 h after study inclusion. The serum samples were kept at −75 °C until they were analyzed at the Virus Diagnostics Laboratory, University of Turku. Serum cytokine levels were determined by 27-plex immunoassay (IL-1β, IL-1rα, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, eotaxin, FGF basic, G-CSF, GM-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, platelet-derived growth factor (PDGF-ββ), MIP-1, RANTES, TNF-α, vascular endothelial growth factor (VEGF)) from Bio-Rad Laboratories, Inc. (California, United States)). According to the manufacturer’s instructions, except that the amount of beads, detection antibodies, and streptavidin-phycoerythrin conjugate were used at 50% of their recommended concentration, which was tested previously as appropriate for the analysis system. The results were analyzed with Bio-Plex Manager 6.0 software. For statistical analyses and calculating the geometric mean cytokine levels, samples under the detection limit were given a value that was the detection limit divided by two. This was done in order to enable the inclusion of negative values (0 values) for geometric mean calculations.

Karhu J., Ala-Kokko T.I., Vuorinen T., Ohtonen P., Julkunen I, & Syrjälä H.T. (2018). Interleukin-5, interleukin-6, interferon induced protein-10, procalcitonin and C-reactive protein among mechanically ventilated severe community-acquired viral and bacterial pneumonia patients. Cytokine, 113, 272-276.

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Western Blot Analysis of Immune Signaling

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Cell lysates of PBECs and tissue lysates of calves after different treatments were prepared by adding RIPA cell lysis buffer (Thermo Fisher Scientific) containing protease and phosphatase inhibitors (Roche Applied Science, Pennsburg, Germany). Total protein content was estimated by bicinchoninic acid analysis (Pierce, Thermo Fisher Scientific) according to the manufacturer’s protocol. Samples were loaded onto polyacrylamide gradient gels (4–20% Tris-HCl, Bio-Rad) and electrotransferred onto polyvinylidene difluoride membranes (Bio-Rad). The membranes were blocked with PBS containing 0.05% Tween-20 (PBST) and 5% milk proteins for 1 h at room temperature and incubated with primary antibodies at 4 °C overnight (NLRP3, 1:1000, PA5-18118; TLR4, 1:1000, PA5-23284, Thermo Fisher Scientific; IL-1β, 1:100, MCA-1658, Bio-Rad; p-p38, 1:1000, #9215; p-ERK1/2, 1:1000, #9101; p-JNK1/2, 1:1000, #9251; p-p65, 1:1000, #3033; β-actin, 1:5000, #4970, Cell Signaling Technology, Beverly, MA), followed by washing blots in PBST. Appropriate horseradish peroxidase-coupled secondary antibodies from Dako (Agilent Technologies, Santa Clara, CA) were applied for 1 h. Membranes were incubated with ECL western blotting substrates (Bio-Rad) prior to obtaining the digital images. Digital images were acquired with the Molecular Imager (Gel DocTM XR, Bio-Rad) and analyzed with Image lab 5.0 (Bio-Rad).

Cai Y., Gilbert M.S., Gerrits W.J., Folkerts G, & Braber S. (2021). Galacto-oligosaccharides alleviate lung inflammation by inhibiting NLRP3 inflammasome activation in vivo and in vitro. Journal of Advanced Research, 39, 305-318.

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Protein Expression Analysis in Macrophages

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RAW264.7 macrophages or lung tissues of different treatment groups. The cells/tissues were lysed with RIPA lysis buffer (Santacruz Biotechnology, Santa Cruz, CA, USA). The proteins were quantified and equally loaded for SDS–PAGE, and after completing the run, PVDF membrane was used to transfer the proteins. The membranes were then blocked with 5% milk (Bio-Rad, Hercules, CA, USA) at room temperature for 1 h. The membranes were then incubated overnight with KLF4, Ang1, iNOS, Cox-2, PCNA, ym1 arg1 and il-1β antibodies (1:1000 in 4% milk, Bio-Rad) at 4 °C overnight and then incubated with the secondary antibodies (1:5000) for 1 h. The bands were visualised with ECL (Bio-Rad). β-actin was used as internal controls.

Almatroodi S.A., Almatroudi A., Alsahli M.A., Aljasir M.A., Syed M.A, & Rahmani A.H. (2020). Epigallocatechin-3-Gallate (EGCG), an Active Compound of Green Tea Attenuates Acute Lung Injury Regulating Macrophage Polarization and Krüpple-Like-Factor 4 (KLF4) Expression. Molecules, 25(12), 2853.

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Multiplex Analysis of Immune Biomarkers

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Biomarkers associated with inflammation, macrophage, and lymphocyte activation were measured by magnetic bead-based multiplex according to the manufacturer’s instructions, including APRIL/TNFSF13, BAFF/TNFSF13B, sCD163, IL-2, IL-1β, IL-4, IL-5, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, IFN-γ, sCD40L, and TNF-α (Bio-Rad, Hercules, CA). Multiplex results were analyzed with Bio-Plex Manager Software (Bio-Rad, Hercules, CA). sCD14 was measured by ELISA according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN) (18 (link)) and analyzed using Tecan’s Magellan™ Software (Tecan US, Morrisvillle, NC).

Baloh C.H., Venturi G.M., Fischer B.M., Sadder L.S., Kim-Chang J.J., Chan C., De Paris K., Yin L., Aldrovandi G.M., Goodenow M.M, & Sleasman J.W. (2023). Biomarkers detected in cord blood predict vaccine responses in young infants. Frontiers in Immunology, 14, 1152538.

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Cytokine and Cell Death Biomarkers ELISA

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Dual antibody sandwich ELISA procedure was adopted to compute the cytokines and cell death biomarkers as per instruction manual provided in ELISA kits procured from Krishgen Biosystems, Mumbai (TNF-α, KB3145, IL-1β, KLR0119, IL-6, KLR0135) and KinesisDX, CA, USA (capase-3, K11-5114 and NF-κB, K11-0288). A typical curve of biomarkers (concentrations standard rat TNF-α 450, 225, 56.25, 28.13, 14.06, 7.03, and 3.51 pg/ml; IL-1β of concentrations 4.8, 2.4, 1.2, 0.6, and 0.3 pg/ml; IL-6 24, 12, 6, 3, and 1.5 pg/ml; NF-κB 12, 6, 3, 1.5, and 0.75 ng/ml; caspase-3 6.4, 3.2, 1.6, 0.8, and 0.4 ng/ml) was plotted to estimate TNF-α (pg/ml), IL-1β (pg/ml), IL-6 (pg/ml), caspase-3 (ng/ml), and NF-κB (ng/ml) in the samples at 450 nm by using ELISA reader (iMARK, BIORAD).

Bhat J.A, & Kumar M. (2022). Neuroprotective Effects of Theobromine in permanent bilateral common carotid artery occlusion rat model of cerebral hypoperfusion. Metabolic brain disease, 37(6).

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Comprehensive Immune Response Evaluation

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Tail DNA was extracted with DirectPCR Lysis Reagent (Viagenbiotech). Lipopolysaccharides from Escherichia coli 055:B5 (#L4005), actinomycin D (#A9415) were purchased from Sigma. Thioglycollate (#236310) was purchased from Becton Dickinson. Taqman and SYBR Green PCR assay kits, GAPDH, MIP1α, MIP-2, CXCL9, CXCL11 and PP2ACα primer/probes were purchased from Applied Biosystems. Mouse TNF-α, IL-6, KC, IL-10, IFNα4 and IFNβ quantitet primers were purchased from Qiagen. Phospho-p38, -p44/42 MAPK, -SAPK/JNK, -c-Jun, -IκBα,-IKKα/β, -NFκB p65, -stat3, -IRF7/IRF3 and non-phospho-IκB antibodies were obtained from Cell Signaling. Anti-PP2A antibody (1D6, #05–421) was purchased from Millipore. Anti-mouse IFNAR-1 antibody (MAR1–5A3) neutralization antibody was purchased from Biolegend. SB 203580 (p38 inhibitor), JNK inhibitor II, Wedelolactone (IKK inhibitor II), FR180204 (ERK inhibitor II) were purchased from Calbiochem. EnzCheck Ser/Thr phosphatase assay kit (#R33700) was purchased from Molecular Probes. Crosslink Magnetic IP/Co-IP kit (#88805) was purchased from Pierce. Mouse TNF-α, IL-10 ELISA Kits were purchased from Invitrogen. Mouse IFN-α ELISA kit was purchased from Life Technologies. A 6-plex kit [IL-1β, IL-10, IL-6, IL-12(p40), IFN-γ, KC and TNF-α] was customized by Bio-Rad.

Sun L., Pham T.T., Cornell T.T., McDonough K.L., McHugh W.M., Blatt N.B., Dahmer M.K, & Shanley T.P. (2016). Myeloid-specific gene deletion of Protein Phosphatase 2A (PP2A) magnifies MyD88- and TRIF-dependent inflammation following endotoxin challenge. Journal of immunology (Baltimore, Md. : 1950), 198(1), 404-416.

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Quantitative RT-PCR Analysis of Immune Markers

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RNA was isolated from cells and tissues using Trizol reagent followed by Direct-zol RNA MiniPrep Plus Kit (Zymo Research) according to manufacturer’s recommendations. Quantitative RT-PCR was performed using one-step RT-PCR kit with SYBR Green (Bio-Rad) and an iQ5 Multicolor Real-Time PCR detection system (Bio-Rad). Primers for RT-PCR (IL-6, IL-1β, TNF, IL-10, Arg-1, IFN-γ, FOXP3, and CD206) were acquired from Bio-Rad and run according to the manufacturer’s protocols. The relative expression level of each mRNA was assessed using the ΔΔCt method and normalized to β-actin/controls.

Thome A.D., Thonhoff J.R., Zhao W., Faridar A., Wang J., Beers D.R, & Appel S.H. (2022). Extracellular Vesicles Derived From Ex Vivo Expanded Regulatory T Cells Modulate In Vitro and In Vivo Inflammation. Frontiers in Immunology, 13, 875825.

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PLGA Nanoparticle-Based Peptide Delivery

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PLGA (Resomer RG503H was purchased from Evonik AG (Germany); PVA (Mowiol 4-88), ethyl acetate and sodium cholate were purchased from Sigma-Aldrich, Germany and was of analytical grade. P2 peptide was purchased from Institute of Medical Immunology, Charité, Berlin, Germany. CFA (Complete Freund’s adjuvant) was purchased from Sigma Aldrich, Germany. Inactivated Mycobacterium tuberculosis particles dissolved in mineral oil was purchased from Difco Laboratories GmbH, Germany. For histology, primary anti-rat antibodies (CD68, MHC-II (OX-6), CD43 (W3/13)) were purchased from BioRad, Germany. For flow cytometry analysis CD16/CD32, CD45 (OX-1, Pecy5) and CD11b (OX-42, PeCy7) were purchased from BD Bioscience, Germany. CD68 (ED1, PE) was purchased from eBioscience, Germany. CD163 (ED2, PE) was purchased from Bio-Rad, Feldkirchen, Germany). Primers for RT-PCR (CD68, CD163, IL-1β, IFN-γ, IL-17, TNF-α) were purchased from Bio-Rad, Germany).

Elahi E., Ali M.E., Zimmermann J., Getts D.R., Müller M, & Lamprecht A. (2022). Immune Modifying Effect of Drug Free Biodegradable Nanoparticles on Disease Course of Experimental Autoimmune Neuritis. Pharmaceutics, 14(11), 2410.

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