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Il 1α

Manufactured by Merck Group
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Sourced in Germany, United States

IL-1α is a recombinant human interleukin-1 alpha protein. It is a pro-inflammatory cytokine that plays a role in the immune response.

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Lab products found in correlation

Il 1β, by Merck Group (5 mentions) Tnf α, by Merck Group (4 mentions) Tnf α, by Cell Signaling Technology (4 mentions) Interleukin 6 (il 6), by Merck Group (3 mentions) Hmgb1, by Novus Biologicals (3 mentions) Lc ms grade solvents, by Merck Group (2 mentions) Milli q h2o purification system, by Merck Group (2 mentions) Il 10, by Merck Group (2 mentions) Human complement c3 elisa kit, by Abcam (2 mentions) Tumor necrosis factor (tnf), by Cell Signaling Technology (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Il 1β, by R&D Systems (2 mentions) Mcp 1, by Merck Group (2 mentions) 3 4 5 dimethyl 2 thiazolyl 2 5 diphenyl 2h tetrazolium bromide mtt, by Merck Group (1 mentions) 2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions) Mmp 2, by Merck Group (1 mentions) Bca assay, by Abcam (1 mentions) Mmp 8, by Merck Group (1 mentions) Mmp 3, by Merck Group (1 mentions) Pro mmp 9, by R&D Systems (1 mentions)

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Human IL-1α (5 citations)

22 protocols using il 1α

1

Antioxidant Activity and Cytotoxicity Evaluation

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6-hydroxy-2,5,7,8-tetramethyl-3,4-dihydrochromene-2-carboxylic acid (Trolox, cat. number 238813), DMSO (cat. number 472301), formic acid (cat. number 00940), ammonium acetate (cat. number 238074), Sephadex™ LH-20 (cat. number GE17-0090), bradykinin acetate (cat. number B3259), tannic acid (cat. number 403040), Folin-Ciocalteu reagent (cat. number 47641), sodium carbonate (cat. number 223530), gallic acid (cat. number G7384), 2′,7′-dichlorofluorescein (cat. number 410217), sodium phosphate dibasic (cat. number S9763), sodium phosphate monobasic monohydrate (cat. number S9638), 2,2′-azobis(2-methylpropionamidine) dihydrochloride (ABAP, cat. number 440914), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS, cat. number A1888), potassium persulfate (cat. number 216224), sodium acetate (cat. number 241245), acetic acid (cat. number 695092), 2,2-diphenyl-1-picrylhydrazyl (DPPH, cat. number D9132), 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT, cat. number M2128), IL-1 α (cat. number SRP3310) and LC–MS grade solvents were purchased from Merck KGaA, Darmstadt, Germany. The peptide LVNEVTEF was custom synthesized by Sigma-Aldrich (Milan, Italy). LC-grade H2O (18 MΩ cm) was prepared with a Milli-Q H2O purification system (Millipore, Bedford, MA, USA).
Baron G., Altomare A., Mol M., Garcia J.L., Correa C., Raucci A., Mancinelli L., Mazzotta S., Fumagalli L., Trunfio G., Tucci L., Lombardo E., Malara D., Janda E., Mollace V., Carini M., Bombardelli E, & Aldini G. (2021). Analytical Profile and Antioxidant and Anti-Inflammatory Activities of the Enriched Polyphenol Fractions Isolated from Bergamot Fruit and Leave. Antioxidants, 10(2), 141.
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2

Cytokine and MMP Quantification in Colon and Kidney

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Proteins were extracted from a 1 cm section of distal colon or kidney as previously described20 (link),37 (link)–39 (link). Concentrations of the innate cytokines IL-1α, IL-1β, IL-6, IL-10 and TNF-α (Merck Millipore, Massachusetts, USA) and matrix metalloproteases (MMP) MMP-2, MMP-3, MMP-8 and Pro-MMP9 (Merck Millipore) were determined by multiplex assay, while TIMP-1 concentrations were determined by ELISA (R&D systems, Minnesota, USA). Kidney pro-MMP-9 concentrations were determined by ELISA (R&D systems). Concentrations were normalized to total protein concentration as determined by a BCA assay (Abcam, UK) as previously described20 (link),38 (link),39 (link).
Dmochowska N., Tieu W., Keller M.D., Hollis C.A., Campaniello M.A., Mavrangelos C., Takhar P, & Hughes P.A. (2020). 89Zr-pro-MMP-9 F(ab′)2 detects colitis induced intestinal and kidney fibrosis. Scientific Reports, 10, 20372.
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3

Multiplexed Cytokine Profiling in Plasma

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Blood from all groups were collected in K2EDTA blood collection tubes and centrifuged at 4 °C for 15 min at 2,000g within 30 min of collection. Plasma was isolated, aliquoted and stored at −80 °C until use. The EMD Millipore’s MILLIPLEX™ MAP Mouse cytokine Magnetic Bead Panel 1 kit was used for the simultaneous quantification of the following analytes: IL-1α, IL-1β, IL-2, IL-5, IL-6, IL-10, IL-13, IL-18, INF-γ and TNF-α (Merck Millipore, Darmstadt, Germany). Luminex method (12 (link)) was used. The method uses a proprietary technique to internally color code microspheres with two fluorescent dyes and to create distinctly colored bead sets of 500 5.6 µm polystyrene microspheres or 80 6.45 µm magnetic microspheres, each of which is coated with a specific capture antibody. After an analyte from a test sample is captured by the bead, a biotinylated detection antibody is introduced. The reaction mixture is then incubated with streptavidin-phycoerythrin (PE) conjugate, the reporter molecule, to complete the reaction on the surface of each microsphere. The Luminex instrument acquires and analyzes data using the LuminexxMAP fluorescent detection method and the LuminexxPONENT™ acquisition software (Thermo Fisher Scientific, Waltham, MA, USA). HMGB1 plasma levels were measured by an ELISA kit specific for rat HMGB1 (ABIN416082) according to the manufacturers' instructions.
Ahmad A., Druzhyna N, & Szabo C. (2016). Delayed treatment with sodium hydrosulfide improves regional blood flow and alleviates cecal ligation and puncture (CLP) - induced septic shock. Shock (Augusta, Ga.), 46(2), 183-193.
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4

Chromatographic Analysis of Polyphenols

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6-Hydroxy-2,5,7,8-tetramethyl-3,4-dihydrochromene-2-carboxylic acid (trolox), (−)-epicatechin, ethyl gallate, protocatechuic acid, Folin–Ciocalteu reagent, sodium carbonate, gallic acid, (+)-catechin, vanillin, hydrochloric acid, potassium chloride, sodium acetate, DMSO, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), IL1α, ethanol, formic acid and LC-MS grade solvents were purchased from Merck KGaA, Darmstadt, Germany. Quercetin 3-galactoside and malvidin 3-glucoside were obtained from Extrasynthese (Genay CEDEX, France). LC-grade H2O (18 MΩ cm) was prepared with a Milli-Q H2O purification system (Millipore, Bedford, MA, USA).
Baron G., Ferrario G., Marinello C., Carini M., Morazzoni P, & Aldini G. (2021). Effect of Extraction Solvent and Temperature on Polyphenol Profiles, Antioxidant and Anti-Inflammatory Effects of Red Grape Skin By-Product. Molecules, 26(18), 5454.
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5

Astrocyte Cytokine-Induced C3 Secretion

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Astrocytes were plated at 2×104 cells/cm2 and treated with 3 ng/ml IL-1α (Sigma), 30ng/ml TNF (Cell Signaling Tech) and 400 ng/ml C1q (MyBioSource) for 24 hours. The media was isolated and spun down to remove debris and the levels of C3 were measured using the Human Complement C3 ELISA Kit (Abcam) as per manufacturer’s instructions.
Tchieu J., Calder E.L., Guttikonda S.R., Gutzwiller E.M., Aromolaran K.A., Steinbeck J.A., Goldstein P.A, & Studer L. (2019). NFIA is a gliogenic switch enabling rapid derivation of functional human astrocytes from pluripotent stem cells. Nature biotechnology, 37(3), 267-275.
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6

NF-κB Reporter Assay in Macrophages

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NF-κB reporter macrophages were seeded in 24-well plates and infected as described above. After 16 h, cells were lysed in 100 μl passive lysis buffer (Promega). Alternatively, 5 × 104 293ET cells were seeded in 24-well plates 24 h prior to use. Transfection with Lipofectamine 2000 (Invitrogen) was performed according to the manufacturer's recommendations with a mixture of 20 ng pRLTK, 50 ng p4kB:Luc, and 250 ng ptCMV-GFP-effector (or pEGFP-N1 control) plasmids. At 24 h after transfection, cells were stimulated overnight (approximately 17 h) with 50 ng/ml TNF-α (Sigma), 10 ng/ml IL-1α (Sigma), 10 ng/ml IL-1β (Sigma), or 4 μg/ml high-molecular-weight (HMW) poly(I·C) (Invitrogen) and subsequently harvested in 100 μl passive lysis buffer (Promega). To autoactivate the NF-κB pathway, cells were transfected with 300 ng pEAKMMP-AU1-TLR4 (or m6pPAC-FLAG-GFP control) vector in addition to the plasmids described above for 24 h prior to harvesting cell lysates. Luciferase activity was measured using a dual luciferase reporter assay system (Promega) and a Tecan Infinite200 PRO plate reader. NF-κB-regulated luciferase activity was normalized to Renilla luciferase intensity and the results are presented relative to either wild-type-infected samples or unstimulated, GFP-expressing control samples.
Günster R.A., Matthews S.A., Holden D.W, & Thurston T.L. (2017). SseK1 and SseK3 Type III Secretion System Effectors Inhibit NF-κB Signaling and Necroptotic Cell Death in Salmonella-Infected Macrophages. Infection and Immunity, 85(3), e00010-17.
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7

Astrocyte Inflammatory Modulation

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Primary human cerebellar astrocytes (ScienCell, USA) were cultured in astrocyte medium (ScienCell, USA). Human astrocytoma cells (U373) were cultured in Dulbecco's modified Eagle's medium (DMEM)/F12 (Life Technologies, USA) containing 10% fetal calf serum (FCS; Life Technologies, USA), and penicillin/streptomycin (50 mg/ml; Life Technologies, USA) in 5% CO2 at 37°C. For treatments, cells were incubated with tumor necrosis factor alpha (TNFα 10 ng/ml; Peprotech, UK) and interferon gamma (IFN‐γ; 10 ng/ml Peprotech, UK), transforming growth factor beta 1 (TGF‐β1; 10 ng/ml, R&D Systems, USA), interleukin 1 alpha (IL‐1α; 3 ng/ml, Sigma‐Aldrich, USA) or Complement component 1q (C1q; 400 ng/ml, Sigma‐Aldrich, USA) for 24 hr.
Kamermans A., Planting K.E., Jalink K., van Horssen J, & de Vries H.E. (2018). Reactive astrocytes in multiple sclerosis impair neuronal outgrowth through TRPM7‐mediated chondroitin sulfate proteoglycan production. Glia, 67(1), 68-77.
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8

Profiling Inflammatory Markers in SARS-CoV-2 Infection

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Patient serum samples and cell culture supernatants from SARS-CoV-2 spike protein transfected cells were analyzed for the presence of secreted IL-6 (Sigma), soluble IL-6R (Life Technologies) IL-1α (Sigma) and HMGB-1 (Novus Biologicals) using ELISA kits following the manufacturer’s instructions. We also analyzed MCP-1 level by ELISA in culture supernatants of LPS stimulated TMNK-1 cells and from SARS-CoV-2 spike expressing A549 cell culture medium in the presence or absence of Tocilizumab (Absolute Antibody).
Patra T., Meyer K., Geerling L., Isbell T.S., Hoft D.F., Brien J., Pinto A.K., Ray R.B, & Ray R. (2020). SARS-CoV-2 spike protein promotes IL-6 trans-signaling by activation of angiotensin II receptor signaling in epithelial cells. PLoS Pathogens, 16(12), e1009128.
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9

Modulating Immune Response to E. coli Infection

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Rats (1–2 month, about 500 g, 10 rats per group) were infected with 109 colony-forming units (CFUs) of E. coli (serotype O55:B5) orally. To simulate the situation of stress-induced LPS accumulation. We set up the group of additional LPS by adding 1 μg/g of LPS (from E. coli serotype O55:B5, Sigma-Aldrich) mixed with E. coli suspension. After 24 h infection, IL-1α, IL-1β, IL-6, intercellular adhesion molecule-1 (ICAM-1) and Tumor Necrosis Factor (TNF-α) from rat serums were detected by the ELISA kits (BD Biosciences) according to the instructions. For further investigating the survival of E. coli infected rats, simultaneous addition of IL-1α (rat recombinant, Sigma-Aldrich) with 10 ng/g for each infected group. Then the ratios of rat survival were recorded. Lastly the E. coli that survived in rat colons were detected by the colony count technique (colony-forming units, CFUs).
Liu X., Zhang H., He S., Mu X., Hu G, & Dong H. (2020). Endothelial-Derived Interleukin-1α Activates Innate Immunity by Promoting the Bactericidal Activity of Transendothelial Neutrophils. Frontiers in Cell and Developmental Biology, 8, 590.
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10

Astrocyte Activation by SAG and Pro-Inflammatory Stimuli

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Cells were transduced with the LV at a multiplicity of infection (MOI) of 10, keeping them in culture for 48, 72 or 96 h. For the treatment with SAG, astrocytes were treated daily with SAG (EMD Millipore Corp., Cat. No. 566660) at 1 μM for 6 consecutive days. As a positive control of A1-reactivity, astrocytes were treated for 24 h with TNF-α (Cell Signaling Technology, Cat. No. 8902), IL-1α (Sigma, Cat. No. SRP6295) and C1q (MyBioSource, Cat. No. MBS143105) at a concentration of 30 ng/ml, 3 ng/ml and 400 ng/ml, respectively. TNF-α and IL-1α were diluted in PBS and stored at − 20 °C and C1q was diluted in H2O and stored at 4 °C. For the autophagy experiments, astrocytes were treated with 50 nM bafilomycin A1 (BafA1, ChemCruz, Cat. No. sc-201550A) for 4 h at 37 °C.
Vicente-Acosta A., Giménez-Cassina A., Díaz-Nido J, & Loria F. (2022). The smoothened agonist SAG reduces mitochondrial dysfunction and neurotoxicity of frataxin-deficient astrocytes. Journal of Neuroinflammation, 19, 93.
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