Human mast cells were cultured from CD34+ progenitor cells and treated with human IgE (Chemicon International, Temecula, Calif) and goat anti-human IgE (1 μg/mL, Sigma) in the presence or absence of diclofenac (10 μmol/L), as described previously.37 (link) Supernatants of the cells were collected and measured for PGD2 and IL-13 with ELISA or stored at −80°C until used as mast cell supernatants for the treatment of ILC2s.
Goat anti human ige
Manufactured by Merck Group
Sourced in United Kingdom
Goat anti-human IgE is a laboratory reagent used to detect and quantify human immunoglobulin E (IgE) in biological samples. It is a polyclonal antibody produced by immunizing goats with human IgE. The antibody binds to human IgE, allowing for its identification and measurement in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin horseradish peroxidase, by R&D Systems (1 mentions)
Horseradish peroxidase, by Merck Group (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by R&D Systems (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions)
Percoll, by Merck Group (1 mentions)
Pipes, by Merck Group (1 mentions)
Zaprinast, by Merck Group (1 mentions)
8 bromo camp, by Merck Group (1 mentions)
8 bromo cgmp, by Merck Group (1 mentions)
Maxisorp microtitre plate, by Thermo Fisher Scientific (1 mentions)
Caffeine, by Merck Group (1 mentions)
Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions)
Goat anti mouse and goat anti rabbit fluorescence dye labelled antibodies, by LI COR (1 mentions)
Rabbit polyclonal antibodies, by Abcam (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Elisa based assay kits for the detection of il 6, by R&D Systems (1 mentions)
4 protocols using goat anti human ige
1
Mast Cell Supernatant Production
2
Quantification of Phl p 7-specific IgG4 and IgE
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ELISA plates (Maxisorp, Nunc) were coated with 5 μg/mL of recombinant Phl p 7 (MRC/Asthma UK Protein Production Facility, King's College London, UK). Plates were blocked with PBS/1% BSA and washed throughout with PBS/0.05% Tween-20. 1 μg/mL purified 102.1F10 IgG4/IgE, negative isotype controls human myeloma IgG4 (Calbiochem), MOv18 IgE34 (link) and positive control Phl p 7 reactive patient serum12 (link) were incubated in triplicate for 2 hours at room temperature, followed by incubation with isotype specific antibodies. IgG4 antibodies were detected with mouse anti-human IgG4 biotin-conjugated antibody (clone G17-4, BD Biosciences), incubated at 1 μg/mL, followed by streptavidin–horseradish peroxidase (R&D Systems). IgE antibodies were detected with polyclonal goat anti-human IgE (Sigma-Aldrich) directly conjugated with horseradish peroxidase using the manufacturer's recommended dilutions. ELISA plates were developed using TMB (R&D Systems) by measuring absorbance at 450 nm.
3
Investigating Immune Cell Signaling
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The following were purchased from the sources indicated; 8-bromo-cAMP, 8-bromo-cGMP, Dimethyl sulfoxide (DMSO), goat anti-human IgE, PIPES (free acid), Percoll, BSA, zaprinast (Sigma, Poole, U.K.);gentamicin, and RPMI 1640 (Gibco BRL, Dundee, U.K.); IL-3 (Peprotech, Rocky Hill, NJ, U.S.A.); Sp-8- CPT-cAMPS and Sp-8-CPT-cGMPS (Biolog Life Science Institute, Bremen, Germany); and ELISA kits for human IL-4 and IL-13 (Mast Diagnostics, Amsterdam, Netherlands).
4
Cellular Signaling Pathway Analysis
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RPMI-1640 medium, foetal calf serum and supplements, caffeine, bovine liver XOD, PGN, goat anti-human-IgE, caffeine metabolites and an ATP luminometric detection kit were purchased from Sigma (Suffolk, UK). AMP/cAMP detection kits were purchased from Promega (Southampton, UK). Maxisorp™ microtitre plates were obtained from Nunc (Roskilde, Denmark) as well as from Oxley Hughes Ltd (London, UK). Mouse monoclonal antibodies to HIF-1α, mTOR and β-actin as well as rabbit polyclonal antibodies against phospho-S2448 mTOR and phospho-T2446 mTOR were from Abcam (Cambridge, UK). Antibodies against phospho-T389 p70 S6 kinase 1 (p70 S6K1), total and phospho-S65 eukaryotic initiation factor 4E binding protein 1 (eIF4E-BP1) antibodies were obtained from Cell Signalling Technology (Danvers, MA USA). Goat anti-mouse and goat anti-rabbit fluorescence dye-labelled antibodies were obtained from Li-Cor (Lincoln, Nebraska USA). ELISA-based assay kits for the detection of IL-6, TNF-α and VEGF were purchased from R&D Systems (Abingdon, UK). All other chemicals were of the highest grade of purity that were commercially available (from either Sigma or Fisher Scientific).
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