Der p 1

Manufactured by Indoor Biotechnologies

Sourced in United States, United Kingdom

Der p 1 is a purified recombinant house dust mite allergen protein produced by Indoor Biotechnologies. It is intended for use in research and development applications.

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Lab products found in correlation

Der p 2, by Indoor Biotechnologies (3 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (2 mentions) Gm csf, by Thermo Fisher Scientific (2 mentions) Rpmi 1640, by Welgene (2 mentions) Trypsin, by Promega (1 mentions) Cli 095, by InvivoGen (1 mentions) Anti tlr2 antibody, by BioLegend (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Aprotinin, by Merck Group (1 mentions) Gm csf, by R&D Systems (1 mentions) Chloroquine, by Merck Group (1 mentions) Lps rs, by Merck Group (1 mentions) Pepinh myd88, by InvivoGen (1 mentions) Pepinh trif, by InvivoGen (1 mentions) Poly 1 c, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) L trp, by Merck Group (1 mentions) Der f 1, by Indoor Biotechnologies (1 mentions)

16 protocols using der p 1

Lung Inflammation in Zfp30 Knockout Mice

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Intratracheal instillation of LPS from E. coli (LIST Biologicals Campbell, CA) into lungs of Zfp30^+/+ and Zfp30^−/− mice was carried out at a dose of 0.3 mg per kg of body weight using previously described methods (Limjunyawong et al. 2015 , Mock et al. 2020 (link)). Bronchoalveolar lavage fluid was collected between 8 and 48 hours after exposure, and differential cell counts in bronchoalveolar lavage fluid were performed. Aliquots of BALF were saved for cytokine quantification. Oropharyngeal aspiration of LPS from E. coli into lungs of Zfp30^+/+ and Zfp30^−/− mice was carried out with 5μg LPS in 40μl PBS.
Dermatophagoides pteronyssinus house dust mite allergen (Der p 1): We used a model of allergic inflammation involving Der p 1 that we previously showed induces predominantly eosinophilic but also neutrophilic inflammation (Kelada et al. 2011 (link)). Zfp30^+/+ and Zfp30^−/− mice were sensitized with 10 μg Der p 1 (Indoor Biotechnologies, Charlottesville, VA) administered through intraperitoneal injection (in 100 μl of PBS) on days 0 and 7 of the experiment, and a 50 μg Der p 1 challenge was administered on day 15 of the experiment (Kelada et al. 2011 (link)). Mice were sacrificed 48–72 hours after challenge, and differential cell counts in bronchoalveolar lavage fluid were performed. Aliquots of BALF were saved for cytokine quantification.

Laudermilk L.T., Tovar A., Homstad A.K., Thomas J.M., McFadden K.M., Tune M.K., Cowley D.O., Mock J.R., Ideraabdullah F, & Kelada S.N. (2020). Baseline and Innate Immune Response Characterization of a Zfp30 Knockout Mouse Strain. Mammalian genome : official journal of the International Mammalian Genome Society, 31(7-8), 205-214.

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Skin Prick Test for Allergies

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Healthy subjects and AD volunteers with no medication were recruited at the Eulji University for blood donation and undergoing skin prick test (Table 1 and Supplementary Table 1). The human blood protocol and associated consent forms were reviewed and approved by the Institutional Review Board (IRB) of the Eulji University. Conventional skin prick test was conducted according to the instructions of the SoluprickR test. Histamine (positive control), DP, DF (ALK Horsholm, Denmark), Der p 1, Der p 2 (INDOOR Biotechnologies, Charlottesville, VA, USA) and Der p 38 were used as test materials. Recombinant allergen proteins (10 μg/mL) were solubilized in phosphate buffered saline (PBS, pH 7.4) before using in this test. A positive result was determined as a ≥1+ (i.e., a wheal ≥3 mm diameter).

Jeon H., Kim G., Kashif A., Hong M.H., Lee J.S., Hong Y., Park B.S., Yang E.J, & Kim I.S. (2021). Pathogenic Mechanism of Der p 38 as a Novel Allergen Homologous to RipA and RipB Proteins in Atopic Dermatitis. Frontiers in Immunology, 12, 646316.

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Allergic Asthma and Sensitization Profiles

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This research was approved by the Medical Ethics Committee of Zhongnan Hospital (approval number: 2017001), and all donors provided written informed consent. There were 76 volunteers who were recruited for participation between January 2017 and May 2017 (32 allergic asthmatic patients, 19 sensitized asymptomatic individuals, and 25 healthy controls (Table 1). All subjects underwent skin prick tests for Dermatophagoides pteronyssinus 1 (Derp1) and common environmental allergens. Allergic asthmatic patients were chosen according to the Global Initiative for Asthma criteria15 : (1) having allergic asthma symptoms, (2) meeting the pulmonary function test criteria of asthma, (3) being monosensitized to Dermatophagoides pteronyssinus 1 (Derp1, Indoor Biotechnologies, Charlottesville, VA, USA) (wheal diameter ≥ 3 mm),16 (link) (4) having no other atopic diseases, and (5) having not used oral or intravenous steroids in the previous 4 weeks. The severity of allergic asthma was assessed on the basis of the Global Initiative for Asthma criteria.15 Sensitized asymptomatic subjects were chosen according to the following criteria: (1) having no allergy symptoms of allergic rhinitis, asthma or other atopic diseases, and (2) being monosensitized to Derp1 (wheal diameter ≥ 3 mm). Healthy controls had no allergic diseases and had negative reactions to Derp1 and common environmental allergens.

Wang W., Wei C., Cheng Z, & Yang J. (2020). Aberrant Th2 Immune Responses Are Associated With a Reduced Frequency of IL-35-Induced Regulatory T Cells After Allergen Exposure in Patients With Allergic Asthma. Allergy, Asthma & Immunology Research, 12(6), 1029-1045.

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Characterization of NAP-Derp1 Interactions

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Purified NAP (10mcg/mL) was reacted with Derp1 (Indoor Biotechnologies, Charlottesville, VA) (5mcg/mL) at in a 2:1 ratio at pH values of 7, 6 and 4. Samples were incubated at 37°C for 1 hour. The post-reaction Der p1 and NAP mixture was visualized on SDS-PAGE gel with Coomassie staining. SDS PAGE was washed in distilled water to remove excess background stain; the gel band corresponding to Der p1 was excised and cut into 1 mm² pieces. The gel pieces were de-stained with 500 mcL of 1:1 acetonitrile and 100 mM ammonium bicarbonate solution for 2–8 hours. Afterwards, 10 mM reductive tris (2-carboxyethyl) phosphine was added and free cysteines alkylated with 55 mM iodoacetamide. Acetonitrile and 100 mM ammonium bicarbonate were used to dehydrate and rehydrate the gel pieces alternatively for three times. Gel pieces were swelled in 50 mM ammonium bicarbonate containing 10 ng/μL trypsin (Promega, Madison, WI), and Glu-C and digested overnight. Peptides were extracted with 60% acetonitrile/5% formic acid and dried in SpeedVac.

Thomas R.G., Rivera Reyes B.M., Gaston B.M., Rivera Acosta N.B., Bederman I.R., Smith L.A., Sutton M.T., Wang B., Hunt J.F, & Bonfield T.L. (2017). Conjugation of nitrated acetaminophen to Der p1 amplifies peripheral blood monocyte response to Der p1. PLoS ONE, 12(12), e0188614.

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Inhibition of Toll-like Receptor 4 Signaling

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RPMI 1640 and fetal bovine serum (FBS) were purchased from Life Technologies Inc. (Gaithersburg, MD). CLI-095, an inhibitor of Toll-like receptor (TLR) 4 (TLR4i) was purchased from Invivogen (San Diego, CA, USA). PKC δ inhibitor (rottlerin) MEK inhibitor (PD98059), and NF-κB inhibitor (BAY-11-7085) were purchased from Calbiochem (San Diego, CA, USA). GM-CSF was purchased from R&D Systems (Minneapolis, MN, USA). FITC-conjugated rabbit anti-mouse IgG, and FITC-conjugated rat anti-rabbit IgG were purchased from Molecular Probes (Eugene, OR, USA). Antibodies against phospho-tyrosine and phospho-ERK1/2 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against ERK2, procaspase 3, and procaspase 9 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-TLR2 antibodies were obtained from Biolegend (San Diego, CA, USA). The protease inhibitors aprotinin and E64 were obtained from Sigma–Aldrich (St. Louis, MO, USA). DP and DF were obtained from the Korea National Arthropods of Medical Importance Resource Bank (Yonsei University, Seoul, Korea) and Cosmo Bio (Tokyo, Japan). Der p1 and Der p2 were purchased from INDOOR biotechnologies (Charlottesville, VA, USA).

Kim E.H., Lee J.S., Lee N.R., Baek S.Y., Kim E.J., Lee S.J, & Kim I.S. (2014). Regulation of Constitutive Neutrophil Apoptosis Due to House Dust Mite Allergen in Normal and Allergic Rhinitis Subjects. PLoS ONE, 9(9), e105814.

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Investigating Inflammatory Responses to TLR Agonists

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Damaged and not-damaged cultures (controls) were stimulated with the TLR agonists poly(I:C) (Sigma-Aldrich) (0.1 µg/mL, 1 µg/mL, 10 µg/mL) and LPS (Sigma-Aldrich) (1 µg/mL, 10 µg/mL, 50 µg/mL, 100 µg/mL) for 60 min; the allergen Der p1 (Indoor Biotechnologies Ltd., Cardiff, UK); and conditioned media from virus-infected epithelial cells. TLR agonists and conditioned media were added either alone or in combination with the TLR inhibitor LPS-RS (Sigma-Aldrich) or the TLR inhibitor chloroquine (Sigma-Aldrich) or the intracellular pathway inhibitor Pepinh-TRIF (InvivoGen, Toulouse, France) or the intracellular pathway inhibitor Pepinh-MyD88 (InvivoGen).

Lewandowska-Polak A., Brauncajs M., Jarzębska M., Pawełczyk M., Kurowski M., Chałubiński M., Makowska J, & Kowalski M.L. (2018). Toll-Like Receptor Agonists Modulate Wound Regeneration in Airway Epithelial Cells. International Journal of Molecular Sciences, 19(8), 2456.

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Murine Model of Allergic Airway Inflammation

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Protocol for sensitization and challenge was based on the previous published method with minor modification (23 (link)). Briefly, mice were injected intraperitoneally (i.p.) on days 0, 3, 5 with HDM extract containing 10 µg of Der p1 (Indoor Biotechnologies, VA, USA) in 200 µL of sterile saline and then challenged intranasally (i.n.) with the same agent on days 10, 12 14, 16, and 17. Control mice were received phosphate-buffered saline (PBS) during the sensitization and challenge phases. Mice were sacrificed 24 hours after the last challenge for airway reactivity measurement.

Zhang X., Zhang M., Li L., Chen W., Zhou W, & Gao J. (2021). IRAK-M knockout promotes allergic airway inflammation, but not airway hyperresponsiveness, in house dust mite-induced experimental asthma model. Journal of Thoracic Disease, 13(3), 1413-1426.

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Dendritic Cell Gene Expression Changes

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To investigate changes in the expression of selected genes from transcriptome sequencing analysis in allergen-treated dendritic cells, a human monocytic cell line, THP-1, was used. THP-1 cells were incubated with 10% fetal bovine serum (FBS), penicillin/streptomycin, 50 µM β-mercaptoethanol (Sigma-Aldrich, Saint Louis, MO, USA), and 2 mM glutamine in RPMI 1640 (WELGENE, Gyeongsan, Korea) at 37 °C. To prepare DCs, THP-1 cells were incubated with IL-4 (100 ng/mL) (Pepro Tech, Rocky Hill, NJ, USA) and GM-CSF (100 ng/mL) (Pepro Tech). At day 2, the cells were replenished with IL-4 and GM-CSF, followed by incubation for 2 days to differentiate into immature DCs. At day 6, Der p 1 (2 μg/mL, Indoor Biotechnologies, Charlottesville, VA, USA) was added to the media for the maturation of DCs, and cells were cultured for up to 6 days. DCs were used to validate selected gene expression changes after Der p 1 stimulation for 2, 4, 8, 12, and 24 h.

Lee K., Han M.R., Yeon J.W., Kim B, & Kim T.H. (2020). Whole Transcriptome Analysis of Myeloid Dendritic Cells Reveals Distinct Genetic Regulation in Patients with Allergies. International Journal of Molecular Sciences, 21(22), 8640.

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Measuring Immunomodulatory Activity in Airway Epithelial Cells

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Calu-3 cells (2 × 10⁵ cells/ml) were seeded in a 24-well plate supplemented with 100 μM l-TRP (Sigma-Aldrich, St. Louis, MO, USA), and grown to confluence for 24 h. Cells were stimulated with various TLR agonists and allergen extracts, including LPS and polyribocytidylic acid (polyI:C) (both from Sigma-Aldrich, St. Louis, MO, USA), synthetic lipoprotein of Mycoplasma salivarium (FSL-1), CpG-ODN2216 (5′-ggGGGACGA:TCGTCgggggg-3′) (both from Invivogen), house dust mite (Greer laboratories) extract and its purified allergen (Der p 1, Indoor Biotechnologies), Bermuda grass pollen, peanut, and German cockroach extracts (all from Greer laboratories). IDO activity was measured by quantification of kynurenines in the culture supernatant using a colorimetric assay as we have described previously [64 ].

Aldajani W.A., Salazar F., Sewell H.F., Knox A, & Ghaemmaghami A.M. (2016). Expression and regulation of immune-modulatory enzyme indoleamine 2,3-dioxygenase (IDO) by human airway epithelial cells and its effect on T cell activation. Oncotarget, 7(36), 57606-57617.

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Allergen Binding to Gold Nanoparticles

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AuNPs (3.5*10¹⁰ NPs/ml, 50 nm, stabilized in 0.1 mM phosphate buffered-saline (PBS), Sigma-Aldrich, St. Louis, MO, USA) were incubated with different concentrations of purified recombinant allergens, i.e. 2.5 μg Bet v 1 (courtesy of C. Ackaert as described in 2014 [41 (link)]), 5 μg Der p 1 (Indoor Biotechnologies INC., Cardiff, United Kingdom) or 3 μg Phl p 5 (Allergopharma GmbH, Reinbek, Germany) as described by O. Cromwell et al. [42 (link)], on a test tube rotator (Snijders, Tilburg, Netherlands) at 4 °C overnight. The excess protein was removed by performing three washing steps of 5 min centrifugation at 18,000 rpm followed by resuspension in the original buffer.

Radauer-Preiml I., Andosch A., Hawranek T., Luetz-Meindl U., Wiederstein M., Horejs-Hoeck J., Himly M., Boyles M, & Duschl A. (2016). Nanoparticle-allergen interactions mediate human allergic responses: protein corona characterization and cellular responses. Particle and Fibre Toxicology, 13, 3.

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