Mouse anti-dinitrophenyl (DNP) IgE and DNP-human serum albumin (HSA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Fexofenadine-HCl (Fexo), a histamine H1 receptor antagonist, was obtained from Korea Pharma (Seoul, Korea). The rabbit polyclonal antibodies specific for phospho-IκB, IKKα/β, ERK1/2, JNK, p38, Akt, β-actin, and total form for IκB, ERK1/2, JNK, p38, and Akt were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Rabbit polyclonal antibodies for phospho-cPLA2α (Ser505), cPLA2α, 5-LO, PLCγ1, IKKα/β lamin B and NF-κB p65 as well as secondary goat anti-rabbit IgG-HRP and rabbit anti-goat IgG-HRP antibodies, total Syk, total LAT, and Bay 61-3606 were purchased from Santa Cruz Biotechnology (Dallas, Texas, USA) and antibodies for phosphotyrosine was purchased from Millipore (Millipore, Billerica, MA, USA). The antibody-reactive bands were visualized with an enhanced chemiluminescence (ECL) system (Pierce Biotechnology, Rockford, IL, USA). The enzyme immnoassay (EIA) kits for PGD2, LTC4, histamine and the antibody for COX-2 were purchased from Cayman Chemicals (Ann Arbor, MI, USA).
Dnp human serum albumin
Manufactured by Merck Group
Sourced in United States
DNP-human serum albumin (HSA) is a chemical compound used in laboratory settings. It is a conjugate of 2,4-dinitrophenol (DNP) and human serum albumin (HSA), a protein found in human blood plasma. This compound is commonly utilized as a reagent or standard in various analytical and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dexamethasone (dex), by Merck Group (4 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (4 mentions)
Mouse anti dinitrophenyl dnp ige, by Merck Group (3 mentions)
Plcγ1, by Santa Cruz Biotechnology (3 mentions)
O phthaldialdehyde, by Merck Group (3 mentions)
α minimum essential medium, by Thermo Fisher Scientific (3 mentions)
Alum adjuvant, by Thermo Fisher Scientific (3 mentions)
Histodenz, by Merck Group (3 mentions)
Antibodies for phosphotyrosine, by Merck Group (2 mentions)
Toluidine blue o, by Merck Group (2 mentions)
Monoclonal dinitrophenol dnp specific ige, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Anti dinitrophenyl dnp ige, by Merck Group (2 mentions)
Erk1 2, by Cell Signaling Technology (2 mentions)
Plcγ1, by Cell Signaling Technology (2 mentions)
Calcium ionophore a23187, by Merck Group (2 mentions)
Evans blue, by Merck Group (2 mentions)
Anti dnp ige, by Merck Group (2 mentions)
Ovalbumin (ova), by Merck Group (2 mentions)
21 protocols using dnp human serum albumin
1
Mouse Anti-DNP IgE Binding Assay
2
Investigating Mast Cell Signaling Pathways
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Dasatinib (Fig. 1A ) was purchased from Selleckchem (Houston, TX, USA) and 4-Amino-5-(4-chlorophenyl)-7-(dimethylethyl) pyrazolo[3,4-d] pyrimidine (PP2) was obtained from Calbiochem (La Jolla, CA, USA). Monoclonal dinitrophenol (DNP)-specific IgE, DNP-Human serum albumin (HSA), Evans blue, toluidine blue O and cetrizine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against the phosphorylated forms of Syk (Tyr32), LAT (Tyr191), PLCγ1 (Tyr783), Akt (Ser473), Erk1/2 (Thr202/Try204), JNK (Thr183/Try185) and p38 (Thr180/Tyr182) were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA). All antibodies were used diluted 1:1000, unless otherwise stated. Antibodies against total forms (dilution fold, 1:100) of Lyn (catalog no. sc-7274), Fyn (catalog no. sc-434), Syk, PLCγ1, Erk1/2, JNK and p38 were obtained from Santa Cruz Biotechnology (Dallas, TX, USA) and LAT (dilution fold, 1:1000) and Actin (dilution fold, 1:5000) antibodies were from EMD Millipore Corporation (Billerica, MA, USA). The media and reagents used for cell culture were from Welgene (Gyeongsangbuk-do, Korea) and Gibco/Life Technologies, Inc (Rockville, MD, USA).
3
Monoclonal Antibody-Based Immunoassay
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Monoclonal mouse anti-DNP IgE (Cat #D8406, clone SPE-7), DNP-human serum albumin (HSA), FITC-dextran (150 kDa) and BMS-345541 were from Sigma-Aldrich (St. Louis, MO). Hilyte Plus 647-conjugated goat anti-mouse IgG (Cat # AS-61057-05-H647) and Hilyte Plus 488-conjugated goat anti-mouse IgG (Cat # 61057-H488) were from Anaspec (Fremont, CA). HRP-conjugated goat anti-mouse (Cat # 115-035-166) and anti-rabbit (Cat # 115-035-003) IgG were from Jackson Immuno Research Laboratories (West Grove, PA). Monoclonal anti-HA (MMS-101R, clone 16B12) was from Covance (Princeton, NJ). Rabbit polyclonal anti SNAP23 antibodies were from Abcam (Cat # ab4114).
4
Mouse PCA Model for Evaluating Anti-Allergic Compounds
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ICR mice were obtained from Koatek (Seoul, Korea) and fed with laboratory feed (Purina, Seoul, Korea) and water ad libitum. Mice were acclimatized in a specific pathogen-free animal facility under conditions of 20–22°C, 40–60% relative humidity, and a 12 h/12 h (light/dark) cycle for at least for 7 d. For PCA, 80 ng of mouse anti-dinitrophenyl (DNP) IgE (Sigma) was intradermally injected into one ear of 7-week old male mice, followed 24 h later by oral administration of EBT (100 and 200 mg/kg) or fexofenadine-HCl, a histanime H1 receptor antagonist (Korea Pharma, Seoul, 50 mg/kg). One hour later, the mice were intravenously challenged with 60 mg of antigen (DNP-human serum albumin (HSA); Sigma) in 200 μl of PBS containing 1% (w/v) Evans blue. The mice were euthanized 1 h after treatment with the antigen, and their ears were removed and dissolved with 400 μl of formamide at 63°C overnight. The amount of dye extravasation was determined colorimetrically at 630 nm.
5
Antibody-Mediated Signaling Pathway Analysis
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Mouse anti-dinitrophenyl (DNP) IgE and DNP-human serum albumin (HSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies used were used: Rabbit polyclonal antibodies specific for phospho-IκB, IKKα/β, ERK1/2, JNK, PLCγ1, p38, β-actin, and total form for IκB, ERK1/2, JNK, p38, and 5-LO were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Rabbit polyclonal antibodies against NF-κB p65, Syk, LAT, PLCγ1, phospho-cPLA2 (Ser505), IKKα/β and lamin B as well as secondary goat anti-rabbit IgG-HRP and rabbit anti-goat IgG-HRP antibodies, total Syk, total LAT, and Bay 61-3606 were purchased from Santa Cruz Biotechnology (Dallas, TX, USA) and antibodies for phosphotyrosine was purchased from Millipore (Millipore, Billerica, MA, USA). The enzyme immnoassay (EIA) kits for PGD2 and LTC4 were purchased from Cayman Chemicals (Ann Arbor, MI, USA).
6
Intradermal IgE-Mediated Allergic Reaction
7
RBL-2H3 Cell Sensitization and miRNA-223 Quantification
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After 24 h of seeding in 6-well tissue culture plates, RBL-2H3 cells were sensitized with 250 ng/mL anti-2,4-dinitrophenyl (DNP) IgE (Sigma-Aldrich) overnight. The cells were then washed twice in Tyrode’s buffer (135 mM NaCl, 5 mM KCl, 1.8 mM CaCl2, 1.0 mM MgCl2, 5.6 mM glucose, 20 mM HEPES, and 1 mg/mL bovine serum albumin at pH 7.4) and triggered with or without 100 ng/mL DNP–human serum albumin (HSA) (Sigma-Aldrich) for 4 h. After stimulation, total RNA was purified from cells using TRIzol Reagent (Invitrogen). To analyze miRNA-223 expression, qRT-PCR was performed using an miRNA reverse transcription kit and TaqMan miRNA assays from Applied Biosystems according to the manufacturer’s instructions.
8
Mast Cell Activation Signaling
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Rabbit anti‐MRGPRX2 antibody (Cat# HPAD5520), Anti‐Dinitrophenyl (DNP) mouse IgE (Cat# D8406), DNP‐Human serum albumin (HSA), Bovine serum albumin, dimethyl sulfoxide, red blood cell lysis buffer, and p‐nitrophenyl‐N‐acetyl‐β‐D glucosaminide were from Sigma‐Aldrich (Darmstadt, Germany). Recombinant murine IL‐3 and SCF were purchased from PeproTech (NJ). The rabbit anti‐PAMP antibody (Cat# orb10050) was obtained from Biorbyt (Cambridge, UK). Alexa Fluor 488‐conjugated avidin, fetal bovine serum, RPMI1640, Pen Strep, and DAPI were from Thermo Fisher (CA). The following reagents and antibodies were from Biolegend (CA): PE anti‐mouse CD117(Cat# 105807), APC anti‐mouse FcεRIα (Cat# 134315), and APC anti‐mouse CD107a (Cat# 121613). PAMP9‐20 was from Sangon Biotech (Shanghai, China).
9
Comparative Asthma Treatment Evaluation
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Hispidulin and dexamethasone were purchased from Sigma-Aldrich (St. Louis, MO, USA). To compare the efficacy of asthma treatment, dexamethasone (Dex) was used as the reference drug. Anti-dinitrophenyl (DNP) IgE, DNP-human serum albumin (HSA), o-phthaldialdehyde, and ovalbumin (OVA) were purchased from Sigma-Aldrich, whereas alum was provided by Thermo Fisher Scientific (Waltham, MA, USA). The enzyme-linked immunosorbent assay (ELISA) kits for mouse IL-4, IL-5, IL-6, and IL-13, for rat tumor necrosis factor (TNF)-α and IL-4, and for eosinophil peroxidase (EPO) were purchased from BD Biosciences, Invitrogen, and Ebioscience (San Diego, CA, USA), respectively.
10
Reagents and Suppliers for Cell Culture
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Dulbecco’s Modified Eagle Medium (DMEM), phosphate-buffered saline (PBS), and penicillin-streptomycin antibiotics (P/S) were supplied by Gibco (Gaithersburg, MD, USA). Fetal bovine serum (FBS) was obtained from GenDEPOT (Barker, TX, USA). Primary antibodies and secondary antibodies were purchased from Abcam (Cambridge, MA, USA). Lipopolysaccharide (LPS), ovalbumin (OVA) from chicken egg, aluminum hydroxide (Alum), anti-dinitrophenyl (DNP) IgE, DNP-human serum albumin (HSA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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