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).
Mouse anti dinitrophenyl dnp ige
Manufactured by Merck Group
Sourced in United States
The Merck Mouse anti-dinitrophenyl (DNP) IgE is a laboratory reagent used for the detection and quantification of IgE antibodies specific to the DNP antigen. It functions as a targeted immunoassay component.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Dnp human serum albumin, by Merck Group (3 mentions)
Horseradish peroxidase conjugated goat anti rabbit secondary antibody, by Cell Signaling Technology (3 mentions)
β actin, by Cell Signaling Technology (3 mentions)
Antibodies for phosphotyrosine, by Merck Group (2 mentions)
Phospho ikkα β, by Cell Signaling Technology (2 mentions)
Phospho iκbα, by Cell Signaling Technology (2 mentions)
Enhanced chemiluminescence ecl western blot detection reagent, by Thermo Fisher Scientific (2 mentions)
Bay 61 3606 reagent, by Santa Cruz Biotechnology (2 mentions)
Ltc4 enzyme linked immunoassay eia kit, by Cayman Chemical (2 mentions)
Antibody for cox 2, by Cayman Chemical (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Pyrrolidine dithiocarbamate, by Merck Group (1 mentions)
Sb203580, by Merck Group (1 mentions)
Imperatorin, by ChromaDex (1 mentions)
Dnp human serum albumin has, by Merck Group (1 mentions)
Phospho plcγ1, by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Cox 2 antibody, by Cayman Chemical (1 mentions)
Pd98059, by Merck Group (1 mentions)
9 protocols using mouse anti dinitrophenyl dnp ige
1
Mouse Anti-DNP IgE Binding Assay
2
Mast Cell Activation Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI-1640, Modified Eagle Medium (MEM), non-essential amino acid solution, and penicillin-streptomycin were acquired from Hyclone (Logan, Utah, USA). Imperatorin was purchased from ChromaDex (Irvine, CA, USA). Mouse anti-dinitrophenyl (DNP) IgE and DNP-human serum albumin (HAS) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The recombinant mouse SCF was purchased from STEMCELL Technologies Inc (Vancouver, BC, Canada). The primary antibodies used in the experiments were as follows: rabbit polyclonal antibodies specific for phospho-ERK1/2, ERK1/2, phosphop38, p38, phospho-JNK, JNK, phospho-PLCγ1, phospho-Akt, Akt, phospho-IκBα, IκBα, phospho-IKKα/β, and β-actin from Cell Signaling Technology, Inc. (Beverly, MA, USA); rabbit polyclonal antibodies for 5-LO, and phospho-cPLA2 from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Specific inhibitors for NF-κB (pyrrolidine dithiocarbamate, PDTC) and MAPKs (SB203580, PD98059, and SP600125) were obtained from Sigma. The horseradish peroxidase-conjugated goat anti-rabbit secondary antibody was from Cell Signaling. 1×RIPA buffer, NE-PER Nuclear Protein Extraction kit, phosphatase/protease inhibitor cocktail, and enhanced chemiluminescence (ECL) detection reagent were from Pierce (Rockford, IL, USA). The enzyme immunoassay (EIA) kits for LTC4 and PGD2, and COX-2 antibody were purchased from Cayman Chemicals (Ann Arbor, MI, USA).
3
Mouse PCA Model for Evaluating Anti-Allergic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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.
4
Antibody-Mediated Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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).
5
Allergic Inflammation Characterization in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BIS, Mouse anti-dinitrophenyl (DNP) IgE, 2,4-dinitrochlorobenzene (DNCB), glycine, cell proliferation Kit II, 4-Nitrophenyl N-acetyl-β-D-glucosaminide (p-NAG), bovine serum albumin (BSA), ketotifen, formamide, Evans Blue, and o-phthaldialdehyde were purchased from Sigma-Aldrich (ST. Louis, MO, USA). The 2,4-dinitrophenyl-human serum albumin (DNP-HSA) and 2,4-dinitrophenyl-bovine serum albumin (DNP-BSA) were from Biosearch technology. The antibodies specific for p-JNK (Thr183/Tyr185), JNK, p-Erk1/2 (Thr202/Tyr204), Erk1/2, p-P38 (Thr180/Tyr182), P38, p-IκBα (Ser32), IκBα, and normal rabbit IgG were from Cell Signaling Technology (Danvers, MA, USA). The antibodies specific for GAPDH were from Proteintech (Chicago, USA). RIPA and BCA protein assay kits were from Beyotime (Beijing, China). Fetal Bovine Serum (FBS), mouse TNF alpha ELISA kit, mouse IL-4 ELISA kit, and mouse IgE ELISA kit were from Thermo Fisher. Recombinant mouse IL-3 and recombinant mouse SCF/c-kit ligand were from RD systems China. ECL Immobilon Western Chemiluminescent HRP Substrate was from Millipore (Billerica, USA). Compound dexamethasone acetate cream was purchased from CR SAN JIU (Shenzhen, China).
6
Mast Cell Signaling and Inflammation Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI1640, fetal bovine serum (FBS) and the enhanced chemiluminescence (ECL) Western blot detection reagent were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, United States). Mouse anti-dinitrophenyl (DNP) IgE was purchased from Sigma Chemicals (St. Louis, MO, United States). DNP-human serum albumin (HSA) was from Biosearch Technologies (Petaluma, CA, United States). The antibodies specific for phospho-ERK1/2, ERK1/2, phospho-p38, p38, phospho-JNK1/2, JNK1/2, phospho-PLCγ, phospho-IκBα, IκBα, phospho-IKKα/β, β-actin, and the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, United States). The antibodies specific for phospho-cPLA2, NF-κB p65, lamin B, LAT, Lyn, Fyn, and Syk, as well as Bay 61-3606 reagent were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, United States). The LTC4 enzyme linked immunoassay (EIA) kit, and the antibody for COX-2 were from Cayman Chemical (Ann Arbor, MI, United States). Histamine ELISA kit was purchased from Demeditec Diagnostics GmbH (Kiel, Germany).
7
Mast Cell Activation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI1640, fetal bovine serum (FBS), and the enhanced chemiluminescence (ECL) Western blot detection reagent were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Mouse anti-dinitrophenyl (DNP) IgE was purchased from Sigma Chemicals (St. Louis, MO, USA). DNP-HSA was from Biosearch Technologies (Petaluma, CA, USA). The antibodies specific for phospho-ERK1/2, ERK1/2, phospho-p38, p38, phospho-PLCγ, phospho-IκB, IκB, phospho-IKKα/β, β-actin, and the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). The antibodies specific for phospho-cPLA2, NF-κB p65, lamin B, Lyn, Fyn, and Syk, as well as Bay 61-3606 reagent were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). The LTC4 enzyme linked immunoassay (EIA) kit, and the antibody for COX-2 were from Cayman Chemical (Ann Arbor, MI, USA). Histamine enzyme linked immunosorbent assay (ELISA) kit was purchased from Demeditec Diagnostics GmbH (Kiel, Germany). AB23A was obtained from Push Bio-technology Co., Ltd (Chengdu, China), of which the purity is ≥98%.
8
Solubilization and Preparation of Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The solvents, drugs and other reagents, including, polyethylene glycol (PEG), dimethylsulphoxide (DMSO), N-formyl methyl-leucyl-phenylalanine (FMLP), zileuton, zymosan, mouse anti-dinitrophenyl (DNP) IgE, dinitrophenyl-conjugated bovine serum albumin (DNP-BSA), foetal bovine serum (FBS), heparin, calcium ionophore A23187, LPS, DTT and glutathione peroxidase (GPx) were obtained from Sigma-Aldrich (St. Louis, MO). All compounds for in vitro experiments were solubilised in DMSO and diluted down in PBS. The final concentration of the solvent did not exceed 0.05%. For in vivo experiments, the compounds were made up in drug vehicle (4% DMSO/67.2% PEG/28.8% PBS).
9
Mast Cell Degranulation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mast cells (stage 3) (1 × 106) were resuspended in 1 mL of PBS and stimulated with c48/80 (Sigma) (5 μg/mL, 60 min, 5% CO2, 37°C). After centrifugation, tryptase concentration in the supernatant was quantified using a Mouse Mast Cell Tryptase (MCT) ELISA Kit (Cusabio). A standard curve was generated using a four-parameter logistic (4-PL) curve fit. Mast cell degranulation (β-hexosaminidase release) was determined according to the protocol of Kuehn et al. (2010) . In brief, G2V-mESCS-MCs and control PMCs were sensitized overnight with 1 μg/mL mouse anti-dinitrophenyl (DNP) IgE (Sigma-Aldrich). Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate. After 90-min incubation at 37°C, the reaction was stopped with glycine buffer (400 mM, pH 10.7). The absorbance was measured at 405 nm (Varioskan Flash microplate reader, Thermo Scientific). The percentage of β-hexosaminidase release was calculated according to the formula: %β-hexosaminidase released = 100 × (supernatant content)/(supernatant + lysate content).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!