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Dnp hsa

Manufactured by Merck Group
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DNP-HSA is a reagent used in biochemical and immunological research applications. It functions as a hapten, which is a small molecule that can be attached to a larger carrier protein, such as human serum albumin (HSA), to generate an immune response. The core function of DNP-HSA is to serve as a model system for studying protein-hapten interactions and immune response mechanisms.

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Lab products found in correlation

Anti dnp ige, by Merck Group (11 mentions) Dnp ige, by Merck Group (8 mentions) Ovalbumin (ova), by Merck Group (7 mentions) 4 nitrophenyl n acetyl β d glucosaminide, by Merck Group (6 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Dnp specific ige antibody, by Merck Group (4 mentions) Lipofectamine, by Thermo Fisher Scientific (4 mentions) Monoclonal dnp specific ige, by Merck Group (4 mentions) Mouse anti dnp ige, by Merck Group (3 mentions) Spe 7, by Merck Group (3 mentions) Oligonucleotides, by Bioneer (3 mentions) Il 33, by Thermo Fisher Scientific (3 mentions) Compound 48 80, by Merck Group (3 mentions) Toluidine blue, by Meilun (3 mentions) Phospho p38, by Cell Signaling Technology (3 mentions) Plus reagent, by Thermo Fisher Scientific (3 mentions) Ionomycin, by Merck Group (3 mentions) Cd117 c kit apc, by Thermo Fisher Scientific (2 mentions) Tnp bsa, by Merck Group (2 mentions) Tnp specific ige antibody, by Merck Group (2 mentions)

Close spelling variants of this product

Dinitrophenyl-labeled human serum albumin (DNP-HSA) 2,4-dinitrophenyl human serum albumin (DNP-HSA), Dinitrophenyl-conjugated humanserum albumin (DNP-HSA; Dinitrophenyl-coupled human serum albumin (DNP-HSA) Dinitrophenyl human serum albumin (DNP-HSA) DNP30-40-conjugated human serum albumin (DNP-HSA;

73 protocols using dnp hsa

1

Murine Allergic Response Assay

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Mice were sensitized by i.p. injection of 1 µg of mouse dinitrophenylated human serum albumin (DNP-HSA)–specific IgE (D8406; Sigma-Aldrich) in 100 µl PBS, and control mice were mock-injected i.p. with 100 µl of PBS. 16 h later, sensitized or non-sensitized control mice were injected i.p. with 250 ng of DNP-HSA (D8406; Sigma-Aldrich), and rectal temperature was measured every 10 min for a period of 60 min.
Tauber M., Basso L., Martin J., Bostan L., Pinto M.M., Thierry G.R., Houmadi R., Serhan N., Loste A., Blériot C., Kamphuis J.B., Grujic M., Kjellén L., Pejler G., Paul C., Dong X., Galli S.J., Reber L.L., Ginhoux F., Bajenoff M., Gentek R, & Gaudenzio N. (2023). Landscape of mast cell populations across organs in mice and humans. The Journal of Experimental Medicine, 220(10), e20230570.
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2

Optimized Immunoassay Protocols for DNP-IgE

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OVA, DNP-IgE, and DNP-HSA were purchased from Sigma Aldrich (MO, United States), with a purity of ≥90.0%. RMPI-1640, fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Gibco (Grand Island, NY/Carlsbad, CA, United States). 2,5-Diphenyltetrazolium bromide (MTT), and DNP-HSA were obtained from Sigma Chemical Co. (St.Louis, MO, United States). Milli-Q water was supplied from a water purification system (Millipore, MA, United States). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG was purchased from Invitrogen (Carlsbad, CA, United States), and Alum Adjuvant (Thermo Scientific, United States). Trigonelline hydrochloride (Sigma-Aldrich, Buchs, Switzerland; -analytical standard) was dissolved in PBS and adjusted to the required concentration.
Zhang W., Zhang Y., Chen S., Zhang H., Yuan M., Xiao L., Lu Y, & Xu H. (2021). Trigonelline, An Alkaloid From Leonurus japonicus Houtt., Suppresses Mast Cell Activation and OVA-Induced Allergic Asthma. Frontiers in Pharmacology, 12, 687970.
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3

Mast Cell Degranulation Assay

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BMDMC culture was established as described elsewhere29 (link). In brief, bone marrow cells were flushed from adult WT mice femurs and tibia, centrifuged and resuspended in RPMI containing 10% fetal bovine serum (FBS), 100 U/mL penicillin, 0.1 mg/mL streptomycin, 25 mmom/L HEPES, 2 mmol/L L-glutamine, 1 mmol/L sodium pyruvate, 1 mmol/L nonessential amino acids, and 1 mmol/L MEM amino acids (all from Sigma) in the presence of 5 ng/ml recombinant mouse IL-3 (Peprotech). A selection process for mature mast cells was for a period of at least 4 weeks with continuous enrichment for non-adherent fraction of mast cell precursors. After this period, cells were phenotypically more than 90% mature as assessed by FACS staining with antibodies against mouse FcεR1-PE (eBioscience, clone MAR-1) and CD117(c-Kit)-APC (eBioscience, clone 2B8; Online Figure I). To induce mast cells degranulation, cells were sensitized overnight with 100 ng/ml mouse anti-DNP-IgE (Sigma), washed and stimulated with 100 ng/ml DNP-HSA (Sigma) for 0.5 or 1 h (fast release) or 6 h (slow release). The efficiency of mediator release was monitored by β-hexaminidase activity in mast cell supernatants (releasates; Online Figure I)30 .
Ponomaryov T., Payne H., Fabritz L., Wagner D.D, & Brill A. (2017). Mast Cells Granular Contents Are Crucial for Deep Vein Thrombosis in Mice. Circulation Research, 121(8), 941-950.
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4

Immunology Protocol: Reagent Preparation

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Oligonucleotides used in this study were commer-cially synthesized by the Bionex Co. (Seoul, Korea). DNP-HSA, TNP-BSA, DNP-specific IgE antibody and TNP-specific IgE antibody were purchased from Sigma. Chemicals used in this study were purchased from Sigma. All other antibodies were purchased from Cell Signaling Co. (Beverly, MA). Anti-mouse and anti-rabbit IgG-horseradish peroxidase-conjugated antibody was purchased from Pierce. Lipofectamine and PlusTM reagent for transfection were purchased from Invitrogen. SOCS1 mimetic peptide (D-T-H-F-R-T-F-R-S-H-S-D-Y-R-R-I) was commercially synthesized by Peptron Company (Daejon, Korea).
Noh K., Kim M., Kim Y., Kim H., Kim H., Byun J., Park Y., Lee H., Lee Y.S., Choe J., Kim Y.M, & Jeoung D. (2017). miR-122-SOCS1-JAK2 axis regulates allergic inflammation and allergic inflammation-promoted cellular interactions. Oncotarget, 8(38), 63155-63176.
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5

In Vivo IgE-Mediated Ear Swelling Assay

Cited 1 time
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PCA was performed as previously described (37 (link)). Briefly, mice were sensitized by intradermal injection of 100 ng anti-DNP IgE mAb (Sigma-Aldrich) into the left ears, whereas the right ears received saline as a control. After 24 h, mice were challenged by retro-orbital injection of 100 μg DNP-HSA (Sigma-Aldrich). The thickness of the ear was measured using a thickness micrometer at baseline and for the indicated time points. Changes in ear thickness were reported relative to baseline.
Abdala-Valencia H., Bryce P.J., Schleimer R.P., Wechsler J.B., Loffredo L.F., Cook-Mills J.M., Hsu C.L, & Berdnikovs S. (2015). Tetraspanin CD151 Is a Negative Regulator of FcεRI-Mediated Mast Cell Activation. Journal of immunology (Baltimore, Md. : 1950), 195(4), 1377-1387.
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6

Allergy-inducing Mouse Model Protocols

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Metronidazole, ampicillin, neomycin sulfate, and vancomycin were obtained from Macklin Biochemical Technology (Shanghai, China). Mouse anti-DNP-IgE monoclonal antibody, DNP-HSA, ovalbumin (OVA), cholera toxin (CT), and aluminum were obtained from Sigma-Aldrich. Mouse IL-4, IL-5, IFN-γ, TNF-α and mMCP-1, histamine, zonluin, and the sIgA ELISA kit were from Dongge Boye (Beijing, China). Mouse ZO-1, Claudin-1, and Occludin polyclonal antibodies were obtained from Lianke Biotechnology (Hangzhou, China). The FastDNA SPIN Kit for soil was from Mpbio (Santa Ana, CA, USA). PicoGrenn dsDNA was from Life Technology. The AXYPREP DNA Gel Extraction Kit was obtained from Axygen Biosciences. The Roche GS FLX Titanium em PCR Kit and Roche GS FLX Sequencing kit were from Roche Applied Science. DNA DL2000Marker was obtained from Takara Bio (Dalian, China).
Zhang Q., Cheng L., Wang J., Hao M, & Che H. (2021). Antibiotic-Induced Gut Microbiota Dysbiosis Damages the Intestinal Barrier, Increasing Food Allergy in Adult Mice. Nutrients, 13(10), 3315.
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7

Measuring Mast Cell Degranulation

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To measure degranulation, BMMCs preloaded with 0.15 μg/ml IgE (SPE-7; Sigma) overnight (37°C, 5% CO2) were washed in sterile PBS, resuspended in Tyrode's buffer (130 mM NaCl, 5 mM KCl, 1.4 mM CaCl2, 1 mM MgCl2, 5.6 mM glucose, and 0.1% BSA in 10 mM HEPES, pH 7.4) and adapted to 37°C. Cells were stimulated with different concentrations of Ag (DNP-HSA; Sigma) for 30 min at 37°C. The degree of degranulation was determined by measuring β-hexosaminidase release (53 (link)).
Gast M., Preisinger C., Nimmerjahn F, & Huber M. (2018). IgG-Independent Co-aggregation of FcεRI and FcγRIIB Results in LYN- and SHIP1-Dependent Tyrosine Phosphorylation of FcγRIIB in Murine Bone Marrow-Derived Mast Cells. Frontiers in Immunology, 9, 1937.
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8

Bacterial Modulation of Mast Cell Degranulation

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BMMC, previously incubated overnight with bacteria at a final concentration of OD600 = 0.2 in the well or without, were sensitized with 1 μg/mL mouse IgE anti‐DNP 2682‐I 24 for 1 h and challenged with DNP‐HSA (Sigma‐Aldrich, St. Louis, MO) for 20 min. β‐hexosaminidase was quantitated in supernatants using an enzymatic assay as described by Malbec et al. 23. The percentage of inhibition of β‐hexosaminidase release in BMMC exposed to bacteria was calculated based on β‐hexosaminidase relase in BMMC not exposed to bacteria (100% activation).
Cassard L., Lalanne A.I., Garault P., Cotillard A., Chervaux C., Wels M., Smokvina T., Daëron M, & Bourdet‐Sicard R. (2016). Individual strains of Lactobacillus paracasei differentially inhibit human basophil and mouse mast cell activation. Immunity, Inflammation and Disease, 4(3), 289-299.
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9

Antibody Production and Cell Transfection

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We purchased oligonucleotides from Bioneer Company (Daejeon Korea). We purchased DNP-HSA (2, 4-dinitrophenyl-human serum albumin) and DNP-specific IgE antibody from Sigma. Anti-mouse and anti-rabbit IgG-horseradish peroxidase-conjugated antibody were purchased from Pierce. We purchased other antibodies from Cell Signaling Co. (Beverly, MA). We purchased in vitro transfection agents (Lipofectamine and PlusTM reagent) from Invitrogen. We purchased mouse recombinant IL-27 protein from R&D systems.
Kwon Y., Kim M., Kim Y., Jeong M.S., Jung H.S, & Jeoung D. (2021). EGR3-HDAC6-IL-27 Axis Mediates Allergic Inflammation and Is Necessary for Tumorigenic Potential of Cancer Cells Enhanced by Allergic Inflammation-Promoted Cellular Interactions. Frontiers in Immunology, 12, 680441.
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10

Calcium Imaging of Mast Cell Activation

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BMMCs were generated from bone marrow cell suspensions in the presence of IL-3,
IL-4, and SCF as described previously. Calcium imaging buffer (CIB; 125 mM
sodium chloride, approximately 3 mM potassium chloride, 2.5 mM, 0.6 mM
MgCl2, 10 mM HEPES, 20 mM glucose, 1.2 mM NaHCO3,
20 mM sucrose, pH 7.4) was used for pre-sensitized BMMCs, while for LAD2 cells,
pre-incubation buffer (4 μM Fluo-3, 0.1% (v/v) pluronic F-127 CIB) together with
LicA (0, 25, 50, 100 μM) was used for 30 min. Then, the cells were washed twice
with CIB and stimulated with DNP-HSA (1 μg/mL, for BMMCs) or streptomycin
(100 ng/mL, for LAD2; Sigma-Aldrich) for the first 10 s. We used a fluorescence
microscope to photograph the cells at ×200 magnification under green light (1 s
per photo). The activation status of MCs was determined by the changes in
fluorescence intensity.
Shu J., Cui X., Liu X., Yu W., Zhang W., Huo X, & Lu C. (2022). Licochalcone A inhibits IgE-mediated allergic reaction through PLC/ERK/STAT3 pathway. International Journal of Immunopathology and Pharmacology, 36, 03946320221135462.
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