Mouse tnf α elisa ready set go

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Mouse TNF-α ELISA Ready-Set-Go is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kit designed for the detection and quantification of mouse tumor necrosis factor-alpha (TNF-α) in biological samples.

Automatically generated - may contain errors

Lab products found in correlation

Mouse il 1β elisa ready set go, by Thermo Fisher Scientific (3 mentions) Mouse il 6 elisa ready set go, by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) H2so4, by Carl Roth (1 mentions) Infinite m200, by Tecan (1 mentions) Nigericin, by InvivoGen (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Flagellin from s typhimurium, by InvivoGen (1 mentions) Z vad fmk, by InvivoGen (1 mentions) Bay11 7082, by InvivoGen (1 mentions) Il 1β enzyme linked immunosorbent assay elisa kit, by R&D Systems (1 mentions) Monosodium urate msu crystal, by InvivoGen (1 mentions) Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions) 3 4 methylenedioxy β nitrostyrene, by Merck Group (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Adenosine triphosphate (atp), by Merck Group (1 mentions) Hemoccult test kit, by Beckman Coulter (1 mentions) Mouse amphiregulin duoset elisa, by R&D Systems (1 mentions) Mouse anti β catenin, by BD (1 mentions) Mab1501r, by Merck Group (1 mentions)

14 protocols using mouse tnf α elisa ready set go

Cytokine ELISA Quantification Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the IL‐6 and TNF‐α ELISAs, mouse IL‐6 ELISA Ready‐SET‐Go! and mouse TNF‐α ELISA Ready‐SET‐go! kits (both eBioscience, San Diego, USA), respectively, were used according to the manufacturer’s protocol. Briefly, the supernatant was collected from each OHSC during every media change (3 times), and these samples were combined before use. ELISA 96‐well plates (Microlon, Greiner, Frickenhausen, Germany) were incubated overnight at 4°C with capture antibody solution in 1× coating buffer. The plate was washed with 1× PBS supplemented with 0.05% tween 20, blocked with 1× assay diluent for 1 h and then washed again. Afterward, cytokine standards and samples were applied and they were incubated for 2 h. After the plates were washed, they were incubated with a detection antibody dilution in 1× assay diluent for 1 h, followed by an additional washing step. Streptavidin‐HRP was incubated for 30 minutes in 1× assay diluent. Before incubation of the plates in tetramethylbenzidine (TMB) substrate, they were washed 2 times. After 15 minutes, the reaction was stopped using 5% H₂SO₄ (Carl Roth, Karlsruhe, Germany). Absorbance was measured at 450 nm using a plate reader (Infinite M200, Tecan, Männedorf, Switzerland).

Richter M., Vidovic N., Biber K., Dolga A., Culmsee C, & Dodel R. (2020). The neuroprotective role of microglial cells against amyloid beta‐mediated toxicity in organotypic hippocampal slice cultures. Brain Pathology, 30(3), 589-602.

+ Open protocol

+ Expand

Inflammasome Activation in Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols

BOT-4-one was provided by Sang-Kyu Ye and Byung-Hak Kim (Seoul National University of College of Medicine, South Korea)^{17 (link)}. Penicillin-streptomycin, fetal bovine serum (FBS), opti-MEM, and RPMI 1640 were purchased from Gibco (Grand Island, NY, USA). IL-1β antibody (AF401)⁸ and IL-1β enzyme-linked immunosorbent assay (ELISA) kit were purchased from R&D Systems (Minneapolis, MN, USA). Antibodies against ASC (AL177)⁸, NLRP3 (Cryo-2)⁸, and caspase-1 (clone Casper-1)⁸ were purchased from Adipogen (San Diego, CA, USA). β-actin (sc-1616)^{43 (link)} and ubiquitin (sc-8017)^{37 (link)} were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Monosodium urate (MSU) crystal, Bay11-7082, silica crystal, nigericin, flagellin from S. typhimurium, poly(dAdT), Pam3CSK4, and zVAD-FMK were purchased from InvivoGen (San Diego, CA, USA). Mouse TNF-α ELISA Ready-SET-Go was purchased from eBioscience (San Diego, CA, USA). A western blot chemiluminescence reagent kit was purchased from Pierce Chemical (Rockford, IL, USA). Polyvinylidene fluoride (PVDF) and nitrocellulose membrane were purchased from Millipore Corporation (Bedford, MA, USA). LPS (Escherichia. coli 026:B6 and 011:B4), 3,4-methylenedioxy-β-nitrostyrene, and ATP were purchased from Sigma-Aldrich (St.Louis, MO, USA). All other chemicals used were of the highest quality among those that are commercially available.

Shim D.W., Shin W.Y., Yu S.H., Kim B.H., Ye S.K., Koppula S., Won H.S., Kang T.B, & Lee K.H. (2017). BOT-4-one attenuates NLRP3 inflammasome activation: NLRP3 alkylation leading to the regulation of its ATPase activity and ubiquitination. Scientific Reports, 7, 15020.

+ Open protocol

+ Expand

Colorectal Cancer Tissue Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

DSS (Molecular weight 36,000-50,000 kDa) was obtained from MP Biomedicals (catalog no. 02160110). Hemoccult Test Kit was obtained from Beckman Coulter. Complete EDTA-free protease inhibitor cocktail was from Roche.
Primary antibodies used for immunohistochemistry (IHC) and western blotting (WB) were: mouse anti-actin (Millipore MAB1501R, diluted 1:1000 for WB), mouse anti-β-catenin (BD Transduction Laboratories #610154, diluted 1:1000 for IHC and 1:2000 for WB), rabbit anti-Ki67 (Abcam Ab15580, diluted 1:500 for IHC), rabbit anti-EGFR phosphorylated at Tyr1068 (Abcam Ab40815, diluted 1:200 for IHC). Rabbit anti-PACS-2 18193 was previously described [21 (link)], and was diluted 1:1000 for WB.
Enzyme-linked immunosorbent assay (ELISA) kits used for analysis of ex vivo colonic tissue culture supernatants were mouse amphiregulin DuoSet ELISA (R&D Systems DY989) and mouse TNF-α ELISA Ready-SET Go! (eBioscience 88-7324).
The primers used for Pacs2 genotyping were 5′-ATG CAT ACC TGC CCT TAG CAG AGG-3′, 5′-TGG AGT CTG AGG TTG AGG CCT TGA G-3′, and 5′-ATG GCG TTA CTT AAG CTA GCT TGC-3′. Primers for Apc^Min/+ genotyping were 5′-GCC ATC CCT TCA CGTTAG-3′, 5′-TTC CAC TTT GGC ATA AGG C-3′ and 5′-TTC TGA GAA AGA CAG AAG TTA-3′. All primers were obtained from TAG Copenhagen.

Dombernowsky S.L., Schwarz J., Samsøe-Petersen J., Albrechtsen R., Jensen K.B., Thomas G, & Kveiborg M. (2017). Loss of PACS-2 delays regeneration in DSS-induced colitis but does not affect the ApcMin model of colorectal cancer. Oncotarget, 8(65), 108303-108315.

+ Open protocol

+ Expand

Isolation and Stimulation of Murine Immune Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolation and culture of lamina propria mononuclear cells (LPMC) and mesenteric lymph node cells (MLN) were carried out as previously reported (Shih et al., 2011 (link)) MLN and LPMC were plated at 2.5 × 10⁵ cells/well in 96 well plates in RPMI 1640 medium (cellgro) with 10% fetal calf serum, 100 U/ml penicillin, 100 mg/ml gentamycin, 2 mM L-glutamine and stimulated with 2 µL/ml LK Activation cocktail for 4 h. The expression of TNF-α, IL-6 and IL-1β in cell supernatants were assayed by ELISA according to manufacturer’s instructions (Mouse TNF-α ELISA Ready-Set-Go, eBioscience, 88–7324-77; Mouse IL-1β ELISA Ready-SET-Go, eBioscience, 88–7013-77; Mouse IL-6 ELISA Ready-SET-Go, eBioscience, 88–7064-77).

Zhang H., Zheng L., Chen J., Fukata M., Ichikawa R., Shih D.Q, & Zhang X. (2017). The protection role of Atg16l1 in CD11c+ dendritic cells in murine colitis. Immunobiology, 222(7), 831-841.

+ Open protocol

+ Expand

CD4+ T Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

As previously reported (Tobar et al., 2006 ). Isolated CD4⁺ T cells were isolated using the EasySep™ Mouse CD4⁺ T Cell Isolation kit (Stem Cell) from spleen from OT-II/RAG2 mice and stained with Cell Trace CSFE (Thermo Fisher Scientific) according to manufacturer’s recommendations. CFSE stained cells were co-cultured (1BMM to 4 CD4+ T cells) with either Atg16l1^f/f or Atg16l1 deficient BMDCs that were exposed to whole OVA or OVA peptide 323–339 for 6 h. Cells from 72 h co-cultures were collected and stained with CD4 GK1.5 (Biolegend) and Live-Dead stain (Fisher Scientific). After gating on live CD4⁺ T cells, CFSE staining was analyzed by flow cytometry to determine the CD4⁺ T cell proliferation. Cells were also stained with MCHII (Biolegend), CD86 (Biolegend) and D80 (Biolegend) to detect the activity of those cells. The expression of TNF-α and IL-1β in cell supernatants were assayed by ELISA according to manufacturer’s instructions (Mouse TNF-α ELISA Ready-Set-Go, eBioscience, 88–7324-77; Mouse IL-1β ELISA Ready-SET-Go, eBioscience, 88–7013-77).

Zhang H., Zheng L., Chen J., Fukata M., Ichikawa R., Shih D.Q, & Zhang X. (2017). The protection role of Atg16l1 in CD11c+ dendritic cells in murine colitis. Immunobiology, 222(7), 831-841.

+ Open protocol

+ Expand

Cytokine Quantification by ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cytokines in cell culture supernatant were measured by sandwich ELISA. Microtiter plates (Costar EIA/RIA plates, Corning Incorporated, USA) were coated with cytokine specific antibody. Interleukin (IL)-10 and tumor necrosis factor α (TNF-α) were measured according to the manufacturer's instruction of OptEIA mouse IL-10 Set (BD Biosciences, USA) and mouse TNF-α ELISA Ready-Set-Go (eBioscience, USA). For IL-6 assay plates were coated with monoclonal rat anti-IL-6 antibodies (BD Biosciences, USA) followed by blocking with 3% low fat milk (IL-6). Recombinant IL-6 (eBioscience, USA) and cell culture supernatant was added into the plates followed by biotinylated rat anti-mouse IL-6 antibody (eBioscience, USA), and streptavidin-HRP (Vector Laboratories, USA). Hydrogen peroxide (Sigma Aldrich, USA) in the presence of chromogenic dye TMB (BioLegend, USA) was used to develop colorimetric reaction. The reaction was stopped with 2 M sulfuric acid (POCH Gliwice, Poland) and the optical density was measured at 450 (570) nm using the microtiter plate reader (PowerWaveX, Bio-Tek Instruments, Winooski, VT).

Strus M., Okoń K., Nowak B., Pilarczyk-Zurek M., Heczko P., Gawda A., Ciszek-Lenda M., Skowron B., Baranowska A, & Marcinkiewicz J. (2016). Distinct effects of Lactobacillus plantarum KL30B and Escherichia coli 3A1 on the induction and development of acute and chronic inflammation. Central-European Journal of Immunology, 40(4), 420-430.

+ Open protocol

+ Expand

Murine BMDM Cytokine Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary bone-marrow-derived macrophages (BMDMs) were obtained from C57BL/6J mice which were maintained under standard specific pathogen-free conditions. BMDMs were differentiated for six days in DMEM (Gibco) containing 10% FCS, Penicillin/Streptomycin (Gibco), 0.1 mM 2-Mercaptoethanol (Gibco) and L929-conditioned medium. On day 6, BMDMs were seeded in 96-well plates at 2.5x10⁴ cells per well in 50 μL culture medium (differentiation medium without L929 supernatants), followed by incubation at 37 °C for 4 h. Cells were incubated with compounds (25 μL of a 4X stock per well) for 2 h followed by stimulation with LPS (ultra-pure LBS-EB from Invivogen, 100 ng/mL final concentration) dispersed in culture medium (25 μL per well) for 18 h. After LPS stimulation, the cell culture medium was removed and clarified by centrifugation for 5 min at 400 × g. Concentrations of TNFα or IL-10 in the supernatants were determined using mouse TNFα ELISA ready-set-go (eBioscience) according to the manufacturer’s instructions. BMDMs were detached with 5 mM EDTA in PBS, transferred to a white plate and viability was assessed using the CellTiter-Glo luminescent cell viability assay (Promega) following to the manufacturer’s instructions. Experiments were performed in biological duplicates and technical triplicates.

Klaeger S., Heinzlmeir S., Wilhelm M., Polzer H., Vick B., Koenig P.A., Reinecke M., Ruprecht B., Petzoldt S., Meng C., Zecha J., Reiter K., Qiao H., Helm D., Koch H., Schoof M., Canevari G., Casale E., Re Depaolini S., Feuchtinger A., Wu Z., Schmidt T., Rueckert L., Becker W., Huenges J., Garz A.K., Gohlke B.O., Zolg D.P., Kayser G., Vooder T., Preissner R., Hahne H., Tonisson N., Kramer K., Goetze K., Bassermann F., Schlegl J., Ehrlich H.C., Aiche S., Walch A., Greif P.A., Schneider S., Felder E.R., Ruland J., Medard G., Jeremias I., Spiekermann K, & Kuster B. (2017). The target landscape of clinical kinase drugs. Science (New York, N.Y.), 358(6367), eaan4368.

+ Open protocol

+ Expand

Measuring Cytokine Responses in Immune Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cytokine measurement in culture supernatants, 1 × 10⁶ total BMDCs, 5 × 10⁴ FACS-purified pDCs, 1 × 10⁵ CAL-1 or 3 × 10⁶ PBMCs were left untreated or pre-treated for 1 h with the SFKs inhibitor PP2 (EMD Millipore), the inhibitor bafetinib (Adooq, Irvine, CA, USA) or the equivalent concentration of DMSO control, followed by 15 h in media in presence or absence of 0.1 μM CpG-B ODN 1668 (Roche), 1 μM CpG-A ODN 2216, 100 μM loxoribine (mouse cells) or 1 μg ml⁻¹ R848 (human cells) (all from InvivoGen, San Diego, CA, USA).
Mouse type I IFN bioactivity was measured with reference to a recombinant mouse IFN-β standard (Research Diagnostics, Concord, MA, USA) using a L-929 cell line transfected with an interferon-sensitive luciferase. Human type I IFN bioactivity was measured with reference to a recombinant human IFN-β standard (InvivoGen) using HEK-Blue IFN-α/β cell line (InvivoGen). Mouse serum IFN-α, mouse and human TNF were measured, respectively, by luminescent ELISA (LumiKine mIFN-α, InvivoGen) or by ELISA (Mouse TNF-α ELISA Ready-SET-Go!, eBioscience, and Human TNF-α ELISA MAX, Biolegend) as described in the manufacturer's protocol.

Dallari S., Macal M., Loureiro M.E., Jo Y., Swanson L., Hesser C., Ghosh P, & Zuniga E.I. (2017). Src family kinases Fyn and Lyn are constitutively activated and mediate plasmacytoid dendritic cell responses. Nature Communications, 8, 14830.

+ Open protocol

+ Expand

Cell Viability and Cytokine Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

All chemicals were of analytical grade. The Dulbecco’s Modified Eagle Medium (DMEM) and RPMI-1640 culture media were purchased from Sigma^® (St. Louis, MO, USA) and the fetal bovine serum from Cultilab^® (Campinas, SP, Brazil). The reagent 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) was purchased from Sigma^® (St. Louis, MO, USA). For the TNF-α release assay, the commercial kit “Mouse TNF-α ELISA Ready-SET-Go!” (EBioscience^®, San Diego, CA, USA) was used.

Dörr J.A., Majolo F., Bortoluzzi L., de Vargas E.Z., Silva J., Pasini M., Stoll S.N., da Rosa R.L., Figueira M.M., Fronza M., Beys-da-Silva W.O., Martins A., Gaspar H., Pedrosa R.P., Laufer S, & Goettert M.I. (2022). Antiulcerogenic Potential of the Ethanolic Extract of Ceiba speciosa (A. St.-Hil.) Ravenna Evaluated by In Vitro and In Vivo Studies. International Journal of Molecular Sciences, 23(24), 15634.

+ Open protocol

+ Expand

Microglia Activation and Cytokine Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Isolated microglia were plated at density of 3 × 10⁴ cells per well on PDL-coated plates. For the assays using inhibitors or antagonists, microglia were pretreated with H-89 (protein kinase A, downstream inhibitor of Gαs; Sigma-Aldrich), UBO-QIC (Gq inhibitor)^{40 (link)}, Y-27632 (Rho-associated coiled-coil-forming kinase; downstream inhibitor of Gα_12/13; Sigma-Aldrich), and JTE907 (CB2-selective antagonist; TOCRIS, Bristol, UK) for 1 hour; with PTX (Gi inhibitor; Wako, Tokyo, Japan) for 6 hours; or with suramin (P2 receptor antagonist; Sigma-Aldrich) for 30 min. After pretreatment, microglia were treated with OAD (Sigma-Aldrich) overnight, and then stimulated with 5 ng/ml of LPS (Sigma-Aldrich) and 0.5 ng/ml of IFN-γ (R&D Systems, MN, USA) for 12 hours. CP-55940 (Sigma-Aldrich), MRS2365 (TOCRIS), LysoPS (P2Y10 agonist^{12 (link)}), P2Y10-selective agonist^{41 (link)} or OAD analogs, including tOAD (Toronto Research Chemicals, Toronto, Canada), oleic acid (Wako, Osaka, Japan), OEtA (Cayman Chemical, MI, USA), OEA (Cayman Chemical), and PEA (Cayman Chemical), were tested as agonists for each GPCR. The concentration of TNF-α in the culture supernatant was quantified by enzyme-linked immunosorbent assay (ELISA) using Mouse TNF-α ELISA Ready-SET-Go (eBioscience, CA, USA).

Kita M., Ano Y., Inoue A, & Aoki J. (2019). Identification of P2Y receptors involved in oleamide-suppressing inflammatory responses in murine microglia and human dendritic cells. Scientific Reports, 9, 3135.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!