Esophageal tissue sections were processed for immunohistochemistry using a primary mAb directed against either TGF-β1 (Santa Cruz Biotechnology Inc, Santa Cruz, CA), IL-5 (R&D), or eotaxin-1 (R&D), as described (16 (link)). Results are expressed as the number of IL-5, eotaxin-1, or TGF-β1 positive cells per mm2 of lamina propria.
Eotaxin 1
Manufactured by R&D Systems
Sourced in United States
Eotaxin-1 is a chemokine that selectively recruits and activates eosinophils. It is involved in the pathogenesis of allergic and inflammatory diseases.
Automatically generated - may contain errors
Lab products found in correlation
Eotaxin 2, by R&D Systems (2 mentions)
Tgf β1, by Santa Cruz Biotechnology (1 mentions)
Unicap system, by Thermo Fisher Scientific (1 mentions)
Luminex performance assay, by R&D Systems (1 mentions)
Eotaxin3, by R&D Systems (1 mentions)
Tissuelyser lt, by Qiagen (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Interleukin 6 (il 6), by BD (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Periostin, by R&D Systems (1 mentions)
Il 17a, by Thermo Fisher Scientific (1 mentions)
Mcp 1, by Thermo Fisher Scientific (1 mentions)
Il 13, by Thermo Fisher Scientific (1 mentions)
Il 25, by Thermo Fisher Scientific (1 mentions)
Il 17f, by Thermo Fisher Scientific (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Il 22, by R&D Systems (1 mentions)
Granzyme b, by R&D Systems (1 mentions)
Interleukin 4 (il 4), by BD (1 mentions)
Softmax pro 3, by Molecular Devices (1 mentions)
7 protocols using eotaxin 1
1
Immunohistochemical Analysis of Esophageal Tissue
2
Biomarker Analysis of Nasal Polyps
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Analysis of biomarker levels in the tissue of nasal polyps was carried out as previously described.5 To create tissue homogenates, the tissue was disrupted at 50 Hz for 2 minutes with the Tissue Lyser LT (Qiagen Benelux, Antwerp, Belgium), and 1 mL 0.9% NaCl in the presence of cOmplete™, an EDTA‐free protease inhibitor (Roche Diagnostics Belgium, Vilvoorde, Belgium), per 0.1 g of tissue was added. Homogenates were centrifuged at 1800 × g for 10 minutes at 4°C. Supernatants were removed and stored at −25°C until analysis.
All supernatants were analyzed for the presence of cytokines, chemokines, ECP, and total IgE. Total IgE and ECP levels were measured using the UniCAP system (Thermo Fisher Scientific, Phadia, Groot‐Bijgaarden, Belgium) according to manufacturer instructions. Cytokines were assayed using the Luminex Performance Assay (IL‐4, IL‐5, IL‐17, tumor necrosis factor alpha [TNF‐α], IL‐10, IL‐1β, IL‐6, and vascular endothelial growth factor) and Luminex Screening Human Assay (IL‐13, IL‐33, TARC [CCL17], eotaxin‐3 [CCL26], eotaxin‐2 [CCL24], eotaxin‐1 [CCL11], and PARC [CCL18]) (R&D Systems Belgium), according to manufacturer instructions.
All supernatants were analyzed for the presence of cytokines, chemokines, ECP, and total IgE. Total IgE and ECP levels were measured using the UniCAP system (Thermo Fisher Scientific, Phadia, Groot‐Bijgaarden, Belgium) according to manufacturer instructions. Cytokines were assayed using the Luminex Performance Assay (IL‐4, IL‐5, IL‐17, tumor necrosis factor alpha [TNF‐α], IL‐10, IL‐1β, IL‐6, and vascular endothelial growth factor) and Luminex Screening Human Assay (IL‐13, IL‐33, TARC [CCL17], eotaxin‐3 [CCL26], eotaxin‐2 [CCL24], eotaxin‐1 [CCL11], and PARC [CCL18]) (R&D Systems Belgium), according to manufacturer instructions.
3
IL-1β and TNF-α Induced Inflammatory Response
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ASM cells were seeded in 6 well plates at 3.2 × 103 cells/ml in 10% foetal bovine serum (FBS) (DSKH, Melbourne, Australia), 1% antibiotics supplemented Dulbecco’s modified Eagle medium (DMEM) (Life Technologies, Carlsbad, CA, USA) for 24 hours, followed by quiescing three days in 0.1% bovine serum albumin (BSA) (Sigma Aldrich, St Louis, Missouri, USA), 1% antibiotics in DMEM prior to being treated with 10 ng/ml IL-1β or 10 ng/ml TNF-α. Supernatant samples were collected over a 24 hour time course (0, 8, 16 and 24 hours) and stored in aliquots at −20 °C until analysis. IL-6 (BD Biosciences, Franklin Lakes, NJ, USA) and eotaxin-1 (R&D Systems, Minneapolis, MN, USA) were detected with an enzyme linked immunosorbent assay (ELISA) kit, according to the manufacturers’ instructions.
4
Quantifying Cytokine and Chemokine Levels in Murine Lungs
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After lung function evaluation, mice were killed under terminal anesthesia (as reported above), and the left lung lobes were removed, immediately frozen in liquid nitrogen, and stored at −80°C. Commercial enzyme immunosorbent assay (ELISA) kits were used for the measurement of cytokine and chemokine proteins in whole-lung homogenates and cell-free supernatants as reported (38 (link)). Briefly, lung tissue was homogenized on ice using a tissue homogeneizer (Omni International, Kennesaw, GA, USA) in 1 ml PBS containing 0.05% Triton X-100 and a protease inhibitor cocktail (Hoffmann-La Roche, Basel, Switzerland). The resulting supernatants were isolated after centrifugation (10,000 × g, 15 min, 4°C). Samples were quantified using commercially available kits [IL-5, IL-13, IL-17 and TNF-α; Duoset, R&D Systems, MN, USA] and eotaxin-1 (R&D Systems), according to the manufacturer’s instructions.
5
Cytokine and Antibody Analysis in Allergy
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The levels of cytokines and other molecules in the BALF were measured by ELISA: IL-4, IL-13, MCP-1, IL-17A, IL-17F and IL-25 (eBioscience) and IL-22, eotaxin 1, eotaxin 2 and periostin (R&D systems) analysis was performed in accordance with manufacturers’ protocols. OVA-specific antibody levels in titrated serum were similarly measured by a sandwich assay, using OVA (plates coated at 20 μg/ml in PBS) and biotin-conjugated-anti-mouse IgG1, IgG2a or IgE (all from BD) antibodies, streptavidin (sv)-HRP and TMB substrate6 8 (link).
6
Cytokine and Chemokine Analysis by ELISA
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Cell culture supernatants and TC fluid were analysed for cytokine and chemokine levels by ELISA according to the manufacturer’s instructions (IL-6, RANTES, granzyme B, eotaxin-1, CCL17: R&D systems, Wiesbaden-Nordenstadt, Germany; IL-4: BD, Heidelberg, Germany; IFN-γ, IL-21: eBioscience). ELISA plates were read and analysed at 450 and 570 nm using a Spectra Max 340pc384 photometer and SOFTmax Pro 3.0 software (Molecular Devices, Sunnyvale, CA, USA).
7
Characterizing Lung Inflammation and Goblet Cells
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