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Anti mouse ige

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The Anti-mouse IgE is a laboratory reagent used for the detection and analysis of mouse immunoglobulin E (IgE) in various research and diagnostic applications. It provides a specific and reliable tool for researchers studying immune responses and allergic reactions in mouse models.

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Lab products found in correlation

Biotin anti mouse ige, by BD (3 mentions) Recombinant ige, by BD (2 mentions) Dnp hsa, by Merck Group (2 mentions) R35 72, by BD (2 mentions) Iscove modified dulbecco media (imdm), by GE Healthcare (1 mentions) Interleukin 6 (il 6), by BD (1 mentions) Mouse tnf α, by BD (1 mentions) Histamine assay kit, by Abnova (1 mentions) Anti mouse igg hrp, by Southern Biotech (1 mentions) Anti mouse iga hrp, by Southern Biotech (1 mentions) Tmb elisa substrate solution, by Thermo Fisher Scientific (1 mentions) Streptavidin hrp, by Rockland Immunochemicals (1 mentions) Anti mouse igm hrp, by Southern Biotech (1 mentions) High binding eia ria 96 well plate, by Corning (1 mentions) Hrp rat anti mouse igg1, by BD (1 mentions) Goat anti mouse ige hrp, by Southern Biotech (1 mentions) Goat anti mouse igg2c hrp, by Southern Biotech (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Compound 48 80, by Merck Group (1 mentions) 2 2 azinobis 3 ethylbenzthiazoline 6 sulfonic acid, by Merck Group (1 mentions)

Close spelling variants of this product

Mouse monoclonal anti‐human IgE antiserum Rat anti-mouse IgE Biotin-labeled anti-mouse IgE detection antibody Biotin-conjugated rat anti-mouse IgE Rat-anti-mouse IgE (R35-72; Biotinylated anti-mouse IgE or IgG1 Mouse anti-human IgE Anti-TNP mouse IgE (clone IgE-3, AP-conjugated mouse anti-human IgE antibody Biotinylated anti-mouse IgE Ab

13 protocols using anti mouse ige

1

Quantifying Murine Antibody Responses

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For serum OVA-specific IgG1 antibody detection, Nunc Maxisorp 96-well plates (Thermo Fisher Scientific) were coated with 10 μg/ml OVA (Grade V, Sigma-Aldrich) O/N at 4°C, subsequently blocked with 1 % BSA and serial dilutions of sera were added (1:1000, 1:2500, 1:5000, 1:10000). Purified mouse IgG1 (BD Biosciences) was used as standard (starting concentration 500 ng/ml). Detection was achieved with biotinylated anti-IgG1 (BD Biosciences) at 0.1 μg/ml. For total IgE detection, the plates were coated with anti-mouse IgE (BD Biosciences) at 2 μg/ml and antibody was detected with biotinylated anti-IgE (BD biosciences) at 0.1 μg/ml. Purified mouse IgE (BD Biosciences) was used as standard (starting concentration 500 ng/ml). For OVA-specific IgE detection, the plates were coated with anti-mouse IgE (BD Biosciences) as above and the antibody was detected by biotin-OVA (Nanocs) at 10 μg/ml. Mouse anti-OVA IgE (Bio-Rad was used as a standard (starting concentration 250 ng/ml). Antibody binding was detected with streptavidin-HRP (BD Biosciences) and developed with TMB substrate set (BD Biosciences).
Agaronyan K., Sharma L., Vaidyanathan B., Glenn K., Yu S., Annicelli C., Wiggen T.D., Penningroth M.R., Hunter R.C., Dela C.S, & Medzhitov R. (2022). Tissue remodeling by an opportunistic pathogen triggers allergic inflammation. Immunity, 55(5), 895-911.e10.
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2

Cytokine and Immunoglobulin Assay Protocols

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Compound 48/80, phorbol 12-myristate 13-acetate (PMA), calcium Ionophore (CI) A23187, avidin peroxidase, and DNCB were purchased from Sigma-Aldrich Chemical Corp., (St. Louis, MO, USA). IMDM was from GE Healthcare Life Science (Little Chalfont, UK) and fetal bovine serum (FBS) was obtained from JR Scientific, Inc., (Woodland, CA, USA). Anti-human IL-4/IL-10, recombinant IL-4/IL-10, biotinylated IL-4/IL-10, anti-mouse IgE, recombinant IgE, and biotinylated IgE were purchased from Pharmingen (San Diego, CA, USA). Human TNF-α, IL-1β, IL-6, IL-8, mouse TNF-α, and IL-6 ELISA kits were purchased from BD Biosciences (San Diego, CA, USA) and the histamine assay kit was obtained from Abnova Corp., (Taipei, Taiwan). Antibodies (Abs) were purchased from Santa Cruz Biotechnology, Inc., (Santa Cruz, CA, USA).
Ok S., Oh S.R., Jung T.S., Jeon S.O., Jung J.W, & Ryu D.S. (2018). Effects of Angelica gigas Nakai as an Anti-Inflammatory Agent in In Vitro and In Vivo Atopic Dermatitis Models. Evidence-based Complementary and Alternative Medicine : eCAM, 2018, 2450712.
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3

Enzyme Immunoassay for Antibody Detection

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High-binding EIA/RIA 96-well plates (Costar) were coated overnight with 10µg/mL BSA-NP32 or BSA-NP5 (Biosearch Technologies Inc) diluted in ELISA coating buffer (28.6mM Na2Co3, 11.9mM NaHCO3, pH 9.6), or 2µg/mL anti-mouse IgE (BD 553413) diluted in PBS. Wells were subsequently washed four times with 0.05% Tween20/PBS, then blocked with 3% BSA/PBS for 2 hours at room temperature. After washing, sera was serially diluted in 1% BSA/PBS, added to wells and incubated for 2 hours at room temperature. Wells were washed and incubated with anti-mouse IgG-HRP (1030-05), anti-mouse IgM-HRP (1021-05), anti-mouse IgA-HRP (1040-05) from Southern Biotech, or anti-mouse IgE-biotin (BD 553419) diluted in 1% BSA/PBS for 2 hours at room temperature. Biotinylated antibodies were further incubated with streptavidin-HRP (Rockland) for 40 minutes at room temperature. After washing, HRP was detected with 1X TMB ELISA Substrate Solution (eBioscience) and color development was stopped with 1M orthophosphoric acid. Plates were analyzed at 450nm with a Biotrak II plate reader.
Bastow C.R., Kara E.E., Tyllis T.S., Vinuesa C.G., McColl S.R, & Comerford I. (2022). TFR Cells Express Functional CCR6 But It Is Dispensable for Their Development and Localization During Splenic Humoral Immune Responses. Frontiers in Immunology, 13, 873586.
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4

Quantification of Allergy-Related Antibodies

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IL-5 and IL-13 were analyzed by ELISA as recommended by the manufacturer (Invitrogen). Total serum IgE was measured with the following reagents: anti-mouse IgE (BD pharmingen, capture antibody; 553413; 1/250), mouse IgE standard (SouthernBiotech; 0114-01), and Goat anti-mouse IgE-HRP (SouthernBiotech detection antibody; 1110-05; 1/16000). HDM and OVA-specific antibodies were analyzed by ELISA in plates that had been coated 3 h at 4 °C overnight with 100 μl HDM (50 μg/ml in PBS) or OVA (50 μg/ml in PBS), with nonspecific binding blocked by incubation for 1 h with 5% BSA. Wells were washed for three times and mouse serum samples diluted in assay diluent were added and incubated for 2 h. Wells were washed three times, then HRP Rat Anti-Mouse IgG1 (BD pharmingen; 559626; 1/250) or Goat Anti-Mouse IgG2c-HRP (SouthernBiotech; 1078-05; 1/4000) was added and incubated for 1 h. Wells were washed five times, and then were incubated with tetramethylbenzidine substrate until the reaction approached saturation. Linear regression analysis or measurement of absorbance was used to determine antibody titers or relative differences.
Huang Y., Zhu L., Cheng S., Dai R., Huang C., Song Y., Peng B., Li X., Wen J., Gong Y., Hu Y., Qian L., Zhu L., Zhang F., Yu L., Yi C., Gu W., Ling Z., Ma L., Tang W., Peng L., Shi G., Zhang Y, & Sun B. (2023). Solar ultraviolet B radiation promotes α-MSH secretion to attenuate the function of ILC2s via the pituitary–lung axis. Nature Communications, 14, 5601.
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5

Immunological Reagents and Cell Culture Protocols

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Compound 48/80, LPS, avidin peroxidase (AP), 2,2′-azinobis (3-ethylbenzthiazoline-6-sulfonic acid (ABTS), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and DNCB were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco’s Modified Eagles Medium (DMEM) was purchased from Gibco BRL (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from JR Scientific, Inc. (Woodland, CA, USA). Anti-mouse TNF-α/IL-6, recombinant TNF-α/IL-6, biotinylated TNF-α/IL-6, anti-mouse IgE, recombinant IgE and biotinylated IgE were purchased from Pharmingen (San Diego, CA, USA). NF-κB, and histone antibodies (Abs) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).
Jung J.W., Kim S.J., Ahn E.M., Oh S.R., Lee H.J., Jeong J.A, & Lee J.Y. (2014). Ribes fasciculatum var. chinense Attenuated Allergic Inflammation In Vivo and In Vitro. Biomolecules & Therapeutics, 22(6), 547-552.
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6

Peanut-specific Antibody Quantification

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PN-specific-IgE, IgG1 and IgG2a in serum was measured as reported previously (50 (link), 51 (link), 59 (link)). Briefly, microtiter plates were coated with peanut extract (sample wells), anti-mouse IgE (BD Biosciences, San Jose, CA, for IgE reference wells), or DNP-HSA (Sigma-Aldrich for IgG2a and IgG1 reference wells) and incubated overnight at 4° C. Subsequently, the plates were blocked with 2% BSA-PBS after washing. Washed plates were incubated with diluted serum samples, mouse IgE (BD Biosciences), anti-DNP-IgG2a, or anti-DNP-IgG1 (Accurate Antibodies, Westbury, NY) overnight at 4°C and later developed by using biotinylated anti-IgE, IgG2a or IgG1 detection antibodies (BD Biosciences), avidin-peroxide, and ABTS substrate (KPL, St Paul, MN).
Yang N., Maskey A.R., Srivastava K., Kim M., Wang Z., Musa I., Shi Y., Gong Y., Fidan O., Wang J., Dunkin D., Chung D., Zhan J., Miao M., Sampson H.A, & Li X.M. (2023). Inhibition of pathologic immunoglobulin E in food allergy by EBF-2 and active compound berberine associated with immunometabolism regulation. Frontiers in Immunology, 14, 1081121.
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7

Quantification of Th2 and Th17 Cytokines and IgE Levels in Asthma Model

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IL-5, IFN-γ (BD Pharmingen), CCL28 (R&D system) ELISA kits and Th17 Milliplex® (Millipore) were used according to the manufacturer’s instructions. For BALF recovered 24 h after the last OVA challenge, IL-5 and IFN-γ were quantified by ELISA. For lung homogenate, concentrations of IL-4, IL-5, IL-13, IL-17a and IL-17f were quantified by Milliplex® in the supernatants obtained after lung tissue was homogenised with an UltraTurax® and centrifuged at 10,000 g for 10 min at 4 °C.
Plasma IgE levels were determined by ELISA. Microtiter plates were coated overnight at 4 °C with the capture antibody, anti-mouse IgE at 2 μg/well in carbonate buffer, pH 9.5 (BD Pharmingen, clone R35-118). OVA-specific IgE levels were measured using ovalbumin-HRP (BUF048, Abdserotec) revealed with TMB reagent set (Pharmingen).
Daubeuf F., Jung F., Douglas G.J., Chevalier E, & Frossard N. (2015). Protective effect of a Protein Epitope Mimetic CCR10 antagonist, POL7085, in a model of allergic eosinophilic airway inflammation. Respiratory Research, 16(1), 77.
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8

HDM-specific IgG and Total IgE ELISA

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For antibody ELISAs, the plate was coated with using 5 μg/ml HDM for specific IgGs. Total IgE in serum was measured using anti-mouse IgE (BD Biosciences, 553413) to coat, mouse IgE (κ, anti-TNP, BD Biosciences, 557079) as standard and biotin anti-mouse IgE (BD Biosciences, 553419) as secondary antibody.
Scibiorek M., Mthembu N., Mangali S., Ngomti A., Ikwegbue P., Brombacher F, & Hadebe S. (2023). IL-4Rα signalling in B cells and T cells play differential roles in acute and chronic atopic dermatitis. Scientific Reports, 13, 144.
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9

Quantifying Mouse IgE and Influenza Antibodies

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Nunc or Immulon 96-well plates were coated overnight at 4°C with anti-mouse IgE (R35–72, BD Pharmingen, catalog no. 553413) antibody. Plates were washed with PBST (0.05% Tween in PBS) and blocked with 1% BSA for 1 hour. Serum samples were incubated at 1:25 dilution along with a mouse IgE standard (C48–2, BD Pharmingen, catalog no. 557080) to determine total IgE or at 1:4 dilution to determine HDM-specific IgE at RT. Plates were washed with PBST and incubated with biotin-conjugated anti-mouse IgE (R35–118, BD Pharmingen, catalog no. 553419) for total IgE or biotin-conjugated HDM (prepared in-house with the EZ-Link Sulfo-NHS-Biotinylation Kit, Thermo Fisher Scientific, catalog no. 21425), for HDM-specific IgE. Plates were washed with PBST and incubated with horseradish peroxidase (HRP) streptavidin (BioLegend, catalog no. 405210) for 30 min at RT. Plates were washed with PBST and developed with trimethylboron substrate (eBioscience, catalog no. 00–4201-56). The reaction was stopped with 2 N H2SO4, and plates were read at 450 nm with a plate reader. For influenza virus–specific ELISA, plates were coated overnight with heat-inactivated X31 virus, serum samples were incubated at 1:1000, and HRP-conjugated anti-mouse IgG light chain (Jackson ImmunoResearch, catalog no. 115–035-174, RRID: AB_2338512) was used for detection.
Castellanos C.A., Ren X., Gonzalez S.L., Li H.K., Schroeder A.W., Liang H.E., Laidlaw B.J., Hu D., Mak A.C., Eng C., Rodríguez-Santana J.R., LeNoir M., Yan Q., Celedón J.C., Burchard E.G., Zamvil S.S., Ishido S., Locksley R.M., Cyster J.G., Huang X, & Shin J.S. (2021). Lymph node–resident dendritic cells drive TH2 cell development involving MARCH1. Science immunology, 6(64), eabh0707.
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10

Basophil Activation Assay in Mice

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To measure basophil activation, blood was taken from the mice on (day 68) and stimulated and analyzed according to Torrero et al.25 In summary, whole blood was collected in heparinized tubes and diluted 1:1 in RPMI 1640 Medium (Gibco, Invitrogen, Carlsbad, CA). Blood was incubated with anti‐mouse IgE at 0.125 µg/mL (R35‐72, BD Biosciences, Franklin Lakes, NJ), PE at 20 µg/mL or medium for 90 min at 37 °C in 5% CO2. Activation was stopped with PBS containing EDTA. After washing, red blood cells were lysed, and cells were fixed using the Whole Blood Lysing Reagents (Beckman Coulter, Fullerton, CA). Cells were incubated with anti‐CD16/CD32 (clone 2.4G2) to block the FcR, then stained with the following fluorescent‐labeled antibodies for 30 min at 4 °C in the dark: anti‐IgE‐FITC (1:100, clone 23G3), anti‐CD49b‐APC (1:200, clone CX5), anti‐CD4‐PE (1:200, clone RM4‐5), and anti‐CD200R‐Percpefluor 710 (1:200, clone OX110), and anti‐CD19‐PE (1:200, clone 6D5) from eBioscience (Breda, the Netherlands). Analysis of the samples was performed on the BD Accuri™ C6 flow cytometer, analysis with BD sampler software (BD Biosciences).
Wagenaar L., Bol‐Schoenmakers M., Giustarini G., Vonk M.M., van Esch B.C., Knippels L.M., Garssen J., Smit J.J, & Pieters R.H. (2018). Dietary Supplementation with Nondigestible Oligosaccharides Reduces Allergic Symptoms and Supports Low Dose Oral Immunotherapy in a Peanut Allergy Mouse Model. Molecular Nutrition & Food Research, 62(20), 1800369.
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