Human IL-33 Antibody Summary
Ser112-Thr270
Accession # O95760
Applications
Human IL-33 Sandwich Immunoassay
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
Cell Proliferation Induced by IL‑33 and Neutralization by Human IL‑33 Antibody. In the presence of sub-optimal amounts of Hamster Anti-Mouse CD3e Monoclonal Antibody (Catalog # MAB484), Recom-binant Human IL-33 (Catalog # 3625-IL) stimulates proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Under these conditions, proliferation elicited by Recombinant Human IL-33 (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3625). The ND50 is typically 0.75-3.0 µg/mL.
IL‑33 in Human Tonsil. IL-33 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL-33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3625) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
IL-33 in Human Tonsil Using Dual RNAscope®ISH and IHC. IL-33 mRNA was detected in formalin-fixed paraffin-embedded tissue sections of human tonsil probed with ACD RNAScope®Probe (Catalog # 400111) and stained using ACD RNAscope®2.5 HD Detection Reagents-Red (top image, Catalog # 32260). Adjacent tissue section was processed for immunohistochemistry using R&D Systems Goat Anti-Human IL-33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3625) at 3 ug/mL with overnight incubation at 4 °C followed by incubation with the Anti-Goat IgG VisUCyte HRP Polymer Antibody (R&D Systems, Catalog # VC004) and DAB chromogen (lower image, yellow-brown). Tissues were counterstained with hematoxylin (blue).
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-33
IL-33, also known as NF-HEV and DVS 27, is a 30 kDa proinflammatory protein that may also regulate gene transcription (1‑3). DVS 27 was identifed as a gene that is up‑regulated in vasospastic cerebral arteries (1). NF-HEV was described as a nuclear factor that is preferentially expressed in the endothelial cells of high endothelial venules relative to endothelial cells from other tissues (2). IL-33 was identified based on sequence and structural homology with IL-1 family cytokines (3). DVS 27, NF-HEV, and IL-33 share 100% amino acid sequence identity. IL-33 is constitutively expressed in smooth muscle and airway epithelia. It is up‑regulated in arterial smooth muscle, dermal fibroblasts, and keratinocytes following IL-1 alpha or IL-1 beta stimulation (1, 3). Similar to IL-1, IL-33 can be cleaved in vitro by caspase-1, generating an N-terminal fragment that is slightly shorter than the C-terminal fragment (3, 4). The N-terminal portion of full length IL-33 contains a predicted bipartite nuclear localization sequence and a homeodomain-like helix-turn-helix DNA binding domain. By immunofluorescence, full length IL-33 localizes to the nucleus in HUVECs and transfectants (2). The C-terminal fragment, corresponding to mature IL-33, binds and triggers signaling through mast cell IL-1 R4/ST2L, a longtime orphan receptor involved in the augmentation of Th2 cell responses (3, 5-7). A ternary signaling complex is formed by the subsequent association of IL-33 and ST2L with IL-1R AcP (8). Stimulation of Th2 polarized lymphocytes with mature IL-33 in vitro induces IL-5 and IL-13 secretion (3). In vivo administration of mature IL-33 promotes increased production of IL-5, IL-13, IgE, and IgA, as well as splenomegaly and inflammatory infiltration of mucosal tissues (3). Full length and mature human IL-33 share 52‑58% aa sequence identity with mouse and rat IL-33. Human IL-33 shares less than 20% aa sequence identity with other IL-1 family proteins.
- Onda, H. et al. (1999) J. Cereb. Blood Flow Metab. 19:1279.
- Baekkevold, E.S. et al. (2003) Am. J. Pathol. 163:69.
- Schmitz, J. et al. (2005) Immunity 23:479.
- Black, R.A. et al. (1989) J. Biol. Chem. 264:5323.
- Xu, D. et al. (1998) J. Exp. Med. 187:787.
- Lohning, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:6930.
- Dinarello, C.A. (2005) Immunity 23:461.
- Chackerian, A.A. et al. (2007) J. Immunol. 179:2551.
Product Datasheets
Citations for Human IL-33 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 10
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The IL-33:ST2 axis is unlikely to play a central fibrogenic role in idiopathic pulmonary fibrosis
Authors: KE Stephenson, J Porte, A Kelly, WA Wallace, CE Huntington, CL Overed-Say, ES Cohen, RG Jenkins, AE John
Respiratory Research, 2023-03-23;24(1):89.
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Oncogenic KRAS-dependent stromal interleukin-33 directs the pancreatic microenvironment to promote tumor growth
Authors: Donahue, KL;Watkoske, HR;Kadiyala, P;Du, W;Brown, K;Scales, MK;Elhossiny, AM;Espinoza, CE;Lasse Opsahl, EL;Griffith, BD;Wen, Y;Sun, L;Velez-Delgado, A;Renollet, NM;Morales, J;Nedzesky, NM;Baliira, RK;Menjivar, RE;Medina-Cabrera, PI;Rao, A;Allen, B;Shi, J;Frankel, TL;Carpenter, ES;Bednar, F;Zhang, Y;Pasca di Magliano, M;
Cancer discovery
Species: Mouse, Transgenic Mouse
Sample Types: Cell Lysates, Whole Tissue
Applications: Immunohistochemistry, Western Blot -
Activator protein transcription factors coordinate human IL-33 expression from noncanonical promoters in chronic airway disease
Authors: Raphael, HE;Hassan, GF;Osorio, OA;Cohen, LS;Payne, MD;Katz-Kiriakos, E;Tata, I;Hicks, J;Byers, DE;Zhang, B;Alexander-Brett, J;
JCI insight
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Symbiotic microbiome Staphylococcus epidermidis restricts IL-33 production in allergic nasal epithelium via limiting the cellular necroptosis
Authors: Jeon, YJ;Gil, CH;Won, J;Jo, A;Kim, HJ;
BMC microbiology
Species: Mouse
Sample Types: Nasal Lavage Fluid
Applications: Western Blot -
Imatinib inhibits pericyte-fibroblast transition and inflammation and promotes axon regeneration by blocking the PDGF-BB/PDGFR? pathway in spinal cord injury
Authors: Yao F, Luo Y, Liu YC et al.
Inflammation and Regeneration
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Organoids capture tissue-specific innate lymphoid cell development in mice and humans
Authors: GM Jowett, E Read, LB Roberts, D Coman, M Vilà Gonzá, T Zabinski, U Niazi, R Reis, TJ Trieu, D Danovi, E Gentleman, L Vallier, MA Curtis, GM Lord, JF Neves
Cell Reports, 2022-08-30;40(9):111281.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Human Melanoma-Associated Mast Cells Display a Distinct Transcriptional Signature Characterized by an Upregulation of the Complement Component 3 That Correlates With Poor Prognosis
Authors: Rajia Bahri, Orsolya Kiss, Ian Prise, Karen M. Garcia-Rodriguez, Haris Atmoko, Julia M. Martínez-Gómez et al.
Frontiers in Immunology
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Fungal mycobiome drives IL-33 secretion and type 2 immunity in pancreatic cancer
Authors: Aftab Alam, Eric Levanduski, Parker Denz, Helena Solleiro Villavicencio, Maulasri Bhatta, Lamees Alhorebi et al.
Cancer Cell
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Nuclear IL-33 regulates cytokinesis and cell motility in normal human epidermal keratinocytes
Authors: H Tsuda, SI Tominaga, M Ohtsuki, M Komine
Journal of dermatological science, 2022-01-25;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Interleukin-33 activates group 2 innate lymphoid cell expansion and modulates endometriosis
Authors: JE Miller, H Lingegowda, LK Symons, O Bougie, SL Young, BA Lessey, M Koti, C Tayade
JCI Insight, 2021-12-08;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IF -
IL-33 Depletion in COVID-19 Lungs
Authors: Rohit Gaurav, Daniel R. Anderson, Stanley J. Radio, Kristina L. Bailey, Bryant R. England, Ted R. Mikuls et al.
Chest
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Pilot Study of Anti-Th2 Immunotherapy for the Treatment of Breast Cancer-Related Upper Extremity Lymphedema
Authors: BJ Mehrara, HJ Park, RP Kataru, J Bromberg, M Coriddi, JE Baik, J Shin, C Li, MR Cavalli, EM Encarnacio, M Lee, KJ Van Zee, E Riedel, JH Dayan
Biology, 2021-09-18;10(9):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
The impact of airborne endotoxin exposure on rheumatoid arthritis-related joint damage, autoantigen expression, autoimmunity, and lung disease
Authors: TR Mikuls, R Gaurav, GM Thiele, BR England, MG Wolfe, BP Shaw, KL Bailey, TA Wyatt, AJ Nelson, MJ Duryee, CD Hunter, D Wang, DJ Romberger, DP Ascherman, JA Poole
International immunopharmacology, 2021-08-27;100(0):108069.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
PPAR? drives IL-33-dependent ILC2 pro-tumoral functions
Authors: G Ercolano, A Gomez-Cade, N Dumauthioz, G Vanoni, M Kreutzfeld, T Wyss, L Michalik, R Loyon, A Ianaro, PC Ho, C Borg, M Kopf, D Merkler, P Krebs, P Romero, S Trabanelli, C Jandus
Nature Communications, 2021-05-05;12(1):2538.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Fibroblast-derived IL-33 facilitates breast cancer metastasis by modifying the immune microenvironment and driving type-2 immunity
Authors: Ophir Shani, Tatiana Vorobyov, Lea Monteran, Dor Lavie, Noam Cohen, Yael Raz et al.
Cancer Research
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IL-33 Alarmin and Its Active Proinflammatory Fragments Are Released in Small Intestine in Celiac Disease
Authors: Federico Perez, Carolina N. Ruera, Emanuel Miculan, Paula Carasi, Karen Dubois-Camacho, Laura Garbi et al.
Frontiers in Immunology
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Symbiotic microbiome Staphylococcus aureus from human nasal mucus modulates IL-33-mediated type 2 immune responses in allergic nasal mucosa
Authors: YJ Jeon, CH Gil, J Won, A Jo, HJ Kim
BMC Microbiol, 2020-10-07;20(1):301.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
High level of interleukin-33 in cancer cells and cancer-associated fibroblasts correlates with good prognosis and suppressed migration in cholangiocarcinoma
Authors: S Yangngam, S Thongchot, A Pongpaibul, K Vaeteewoot, S Pinlaor, P Thuwajit, S Okada, MA Hermoso, C Thuwajit
J Cancer, 2020-09-23;11(22):6571-6581.
Species: Human
Sample Types: Cell Lysates, Whole Tissue
Applications: IHC, Western Blot -
Interleukin-33 Signaling Controls the Development of Iron-Recycling Macrophages
Authors: Yuning Lu, Gemma Basatemur, Ian C. Scott, Davide Chiarugi, Marc Clement, James Harrison et al.
Immunity
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Myristoylation of LMCD1 Leads to Its Species-Specific Derepression of E2F1 and NFATc1 in the Modulation of CDC6 and IL-33 Expression During Development of Vascular Lesions
Authors: Suresh Govatati, Prahalathan Pichavaram, Jagadeesh Janjanam, Liang Guo, Renu Virmani, Gadiparthi N. Rao
Arteriosclerosis, Thrombosis, and Vascular Biology
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ILC2s amplify PD-1 blockade by activating tissue-specific cancer immunity
Authors: JA Moral, J Leung, LA Rojas, J Ruan, J Zhao, Z Sethna, A Ramnarain, B Gasmi, M Gururajan, D Redmond, G Askan, U Bhanot, E Elyada, Y Park, DA Tuveson, M Gönen, SD Leach, JD Wolchok, RP DeMatteo, T Merghoub, VP Balachandr
Nature, 2020-02-19;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC -
IL-33/ST2 axis promotes glioblastoma cell invasion by accumulating tenascin-C
Authors: JF Zhang, T Tao, K Wang, GX Zhang, Y Yan, HR Lin, Y Li, MW Guan, JJ Yu, XD Wang
Sci Rep, 2019-12-30;9(1):20276.
Species: Human
Sample Types: Tissue
Applications: IHC -
Identification of TNFR2 and IL-33 as therapeutic targets in localized fibrosis
Authors: D Izadi, TB Layton, L Williams, F McCann, M Cabrita, AI Espirito S, W Xie, M Fritzsche, H Colin-York, M Feldmann, KS Midwood, J Nanchahal
Sci Adv, 2019-12-04;5(12):eaay0370.
Species: Human
Sample Types: Whole Cells
Applications: ICC -
The reciprocal interaction between tumor cells and activated fibroblasts mediated by TNF-alpha/IL-33/ST2L signaling promotes gastric cancer metastasis
Authors: Q Zhou, X Wu, X Wang, Z Yu, T Pan, Z Li, X Chang, Z Jin, J Li, Z Zhu, B Liu, L Su
Oncogene, 2019-10-28;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture -
Interleukin 33/ST2 Axis Components Are Associated to Desmoplasia, a Metastasis-Related Factor in Colorectal Cancer
Authors: Glauben Landskron, Marjorie De la Fuente López, Karen Dubois-Camacho, David Díaz-Jiménez, Octavio Orellana-Serradell, Diego Romero et al.
Frontiers in Immunology
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IL-33 drives group 2 innate lymphoid cell-mediated protection during Clostridium difficile infection
Authors: AL Frisbee, MM Saleh, MK Young, JL Leslie, ME Simpson, MM Abhyankar, CA Cowardin, JZ Ma, P Pramoonjag, SD Turner, AP Liou, EL Buonomo, WA Petri
Nat Commun, 2019-06-20;10(1):2712.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
NFATc1-E2F1-LMCD1–Mediated IL-33 Expression by Thrombin Is Required for Injury-Induced Neointima Formation
Authors: Suresh Govatati, Prahalathan Pichavaram, Jagadeesh Janjanam, Baolin Zhang, Nikhlesh K. Singh, Arul M. Mani et al.
Arteriosclerosis, Thrombosis, and Vascular Biology
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Hedgehog Signaling Modulates Interleukin‐33‐Dependent Extrahepatic Bile Duct Cell Proliferation in Mice
Authors: Nataliya Razumilava, Junya Shiota, Nureen H. Mohamad Zaki, Ramon Ocadiz‐Ruiz, Christine M. Cieslak, Kais Zakharia et al.
Hepatology Communications
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Neoplastic Transformation of the Peribiliary Stem Cell Niche in Cholangiocarcinoma Arisen in Primary Sclerosing Cholangitis
Authors: Guido Carpino, Vincenzo Cardinale, Trine Folseraas, Diletta Overi, Krzysztof Grzyb, Daniele Costantini et al.
Hepatology
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Cellular and clinicopathological features of the IL-33/ST2 axis in human esophageal squamous cell carcinomas
Authors: G Cui, J Ren, G Xu, Z Li, W Zheng, A Yuan
Cancer Cell Int., 2018-12-11;18(0):203.
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
Dysregulated invertebrate tropomyosin-dectin-1 interaction confers susceptibility to allergic diseases
Authors: N Gour, S Lajoie, U Smole, M White, D Hu, P Goddard, S Huntsman, C Eng, A Mak, S Oh, JH Kim, A Sharma, S Plante, IH Salem, Y Resch, X Xiao, N Yao, A Singh, S Vrtala, J Chakir, EG Burchard, AP Lane, M Wills-Karp
Sci Immunol, 2018-02-23;3(20):.
Species: Human, Mouse
Sample Types: BALF, Cell Culture Supernates
Applications: ELISA Capture -
HpARI Protein Secreted by a Helminth Parasite Suppresses Interleukin-33
Authors: Megan Osbourn, Dinesh C. Soares, Francesco Vacca, E. Suzanne Cohen, Ian C. Scott, William F. Gregory et al.
Immunity
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Epithelial-derived IL-33 promotes intestinal tumorigenesis in Apc Min/+ mice
Authors: Zhengxiang He, Lili Chen, Fabricio O. Souto, Claudia Canasto-Chibuque, Gerold Bongers, Madhura Deshpande et al.
Scientific Reports
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IL‐10‐producing, ST2‐expressing Foxp3 + T cells in multiple sclerosis brain lesions
Authors: Stephanie Elizabeth Johanna Zandee, Richard Anthony O'Connor, Iris Mair, Melanie Dawn Leech, Anna Williams, Stephen Mark Anderton
Immunology & Cell Biology
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The ”Alarmins” HMBG1 and IL-33 Downregulate Structural Skin Barrier Proteins and Impair Epidermal Growth
Authors: Uffe Nygaard, Ellen H van den Bogaard, Hanna Niehues, Malene Hvid, Mette Deleuran, Claus Johansen et al.
Acta Dermato Venereologica
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Interleukin-33 and RANK-L Interplay in the Alveolar Bone Loss Associated to Periodontitis
PLoS ONE, 2016-12-19;11(12):e0168080.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-P -
Loss of interleukin 33 expression in colonic crypts - a potential marker for disease remission in ulcerative colitis
Authors: Mona Dixon Gundersen, Rasmus Goll, Johanna Hol, Trine Olsen, Renathe Rismo, Sveinung W. Sørbye et al.
Scientific Reports
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TLR2 Activation Limits Rhinovirus-Stimulated CXCL-10 by Attenuating IRAK-1-Dependent IL-33 Receptor Signaling in Human Bronchial Epithelial Cells
J Immunol, 2016-08-08;0(0):.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Interleukin-33 in human gliomas: Expression and prognostic significance
Authors: DOROTHEE GRAMATZKI, KARL FREI, GIERI CATHOMAS, HOLGER MOCH, MICHAEL WELLER, KIRSTEN DIANA MERTZ
Oncology Letters
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Role of IL-33 expression in oncogenesis and development of human hepatocellular carcinoma
Authors: YAN YANG, JUN-BIN WANG, YU-MEI LI, Y U ZHAO, RUI WANG, QIONG WU et al.
Oncology Letters
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The IL-33/ST2 pathway contributes to intestinal tumorigenesis in humans and mice
Authors: Kirsten D. Mertz, Lukas F. Mager, Marie-Hélène Wasmer, Thore Thiesler, Viktor H. Koelzer, Giulia Ruzzante et al.
OncoImmunology
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IL-33 Signaling Protects from Murine Oxazolone Colitis by Supporting Intestinal Epithelial Function
Authors: Amanda Waddell, Jefferson E. Vallance, Preston D. Moore, Amy T. Hummel, David Wu, Shiva K. Shanmukhappa et al.
Inflammatory Bowel Diseases
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Oxidation of the alarmin IL-33 regulates ST2-dependent inflammation
Authors: E. Suzanne Cohen, Ian C. Scott, Jayesh B. Majithiya, Laura Rapley, Benjamin P. Kemp, Elizabeth England et al.
Nature Communications
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IL-33 signaling contributes to the pathogenesis of myeloproliferative neoplasms.
Authors: Mager L, Riether C, Schurch C, Banz Y, Wasmer M, Stuber R, Theocharides A, Li X, Xia Y, Saito H, Nakae S, Baerlocher G, Manz M, McCoy K, Macpherson A, Ochsenbein A, Beutler B, Krebs P
J Clin Invest, 2015-05-26;125(7):2579-91.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Epidermal Expression and Regulation of Interleukin-33 during Homeostasis and Inflammation: Strong Species Differences.
Authors: Sundnes O, Pietka W, Loos T, Sponheim J, Rankin A, Pflanz S, Bertelsen V, Sitek J, Hol J, Haraldsen G, Khnykin D
J Invest Dermatol, 2015-03-04;135(7):1771-80.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Neutralisation of the interleukin-33/ST2 pathway ameliorates experimental colitis through enhancement of mucosal healing in mice
Authors: Mamdouh A K Sedhom, Mélanie Pichery, Jenna R Murdoch, Benoit Foligné, Nathalie Ortega, Sylvain Normand et al.
Gut
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The role of IL-33 and its receptor ST2 in human nasal epithelium with allergic rhinitis.
Authors: Kamekura R, Kojima T, Takano K, Go M, Sawada N, Himi T
Clin. Exp. Allergy, 2012-02-01;42(2):218-28.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization -
Distinct serum and synovial fluid interleukin (IL)-33 levels in rheumatoid arthritis, psoriatic arthritis and osteoarthritis
Authors: Dominique Talabot-Ayer, Thomas McKee, Patrizia Gindre, Sylvette Bas, Dominique L. Baeten, Cem Gabay et al.
Joint Bone Spine
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Measurement of interleukin-33 (IL-33) and IL-33 receptors (sST2 and ST2L) in patients with rheumatoid arthritis.
Authors: Hong YS, Moon SJ, Moon S, Joo YB, Joo Y, Jeon CH, Jeon C, Cho ML, Ju JH, Oh HJ, Heo YJ, Heo Y, Park SH, Kim HY, Min JK
J Korean Med Sci, 2011-09-01;26(9):1132-9.
Species: Human
Sample Types: Synovial Fluid, Whole Tissue
Applications: ELISA Development, IHC-P -
The immune-modulating cytokine and endogenous Alarmin interleukin-33 is upregulated in skin exposed to inflammatory UVB radiation.
Authors: Byrne SN, Beaugie C, O'Sullivan C, Leighton S, Halliday GM
Am. J. Pathol., 2011-05-13;179(1):211-22.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-Fr -
Expression of interleukin 1-like cytokine interleukin 33 and its receptor complex (ST2L and IL1RAcP) in human pancreatic myofibroblasts.
Authors: Nishida A, Andoh A, Imaeda H, Inatomi O, Shiomi H, Fujiyama Y
Gut, 2009-12-08;59(4):531-41.
Species: Human
Sample Types: Cell Lysates, Whole Tissue
Applications: IHC-P, Western Blot -
Inhibition of interleukin-33 signaling attenuates the severity of experimental arthritis.
Authors: Palmer G, Talabot-Ayer D, Lamacchia C, Toy D, Seemayer CA, Viatte S, Finckh A, Smith DE, Gabay C
Arthritis Rheum., 2009-03-01;60(3):738-49.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Eosinophils regulate intra-adipose axonal plasticity
Authors: Meng X, Qian X, Ding X et al.
Proceedings of the National Academy of Sciences of the United States of America
FAQs
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Does the Human IL-33 Antibody, Catalog # AF3625, detect full length IL-33 or the cleaved forms?
The immunogen sequence that this antibody recognizes is Ser112-Thr270 from Accession # O95760. This seqeunce corresponds to the cleaved form and is included in the full length sequence. Therefore, we would expect that this antibody detects both the full length and cleaved forms.
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We used this antibody in an in-house ELISA along with mAb (3625) and protein (3625-IL-010) to quantify IL-33 in human serum and plasma. This combination could not detect IL-33 in our samples but generated a good standard curve.
Staining was done on human airways after citrate antigen retrieval. OCT sections were used
Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287
Used at 10ug/ml.
Briefly, frozen brain sections were fixed in 4% PFA
(Fisher Scientific), followed by antigen retrieval using heating in acid
citric buffer (Vector, Burlingame, CA, USA). Endogenous avidin-biotin was blocked for 15 min (Vector).
Sections were incubated with 10% horse serum in PBS (Biosera, Boussens, France) and Fc Receptor
Blocking Solution was added (Human TruStain FcX Biolegend,
London, UK). Primary antibodies were added overnight at 4 °C.
IL-33 was detected with donkey anti-goat-biotin (ab6578,
Abcam), followed by streptavidin-HRP and visualised with DAB. Section was counterstained with Hematoxylin.
AF3625 was used as both the capture and the detection antibody for the sandwich ELISA for IL-33. The immunoassay standard was 3625-IL. Assay had sensitivity of ~1pg/ml. Many healthy human samples were not detectable.