12-7052-81

antibody from Invitrogen Antibodies

Targeting: IL5 EDF, IL-5, TRF

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

12-7052-81

Provider

Invitrogen Antibodies

Product name

IL-5 Monoclonal Antibody (TRFK5), PE, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The TRFK5 antibody reacts with mouse and human interleukin-5 (IL-5). The TRFK5 antibody is a neutralizing antibody. mouse IL-5 is a disulfide-linked homodimer, containing two 113 amino acid peptides; these resolve as 32-34 kDa bands in SDS-PAGE. IL-5 is produced by T cells and has been known as eosinophil-differentiating factor (EDF), B cell growth factor II (BCGFII), and T cell-replacing factor (TRF). IL-5 induces eosinophil differentiation and promotes eosinophil survival and activation. In mice, IL-5 has been shown to stimulate B cell proliferation and antibody production. Applications Reported: This TRFK5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This TRFK5 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Rat

Conjugate

Yellow dye

Isotype

IgG

Antibody clone number

TRFK5

Vial size

50 µg

Concentration

0.2 mg/mL

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

Lysosomal Acid Lipase Is Required for Donor T Cells to Induce Graft-versus-Host Disease.

Nguyen HD, Ticer T, Bastian D, Kuril S, Li H, Du H, Yan C, Yu XZ
Cell reports 2020 Oct 27;33(4):108316

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CD4(+) T Cells Cross-Reactive with Dengue and Zika Viruses Protect against Zika Virus Infection.

Wen J, Wang YT, Valentine KM, Dos Santos Alves RP, Xu Z, Regla-Nava JA, Ngono AE, Young MP, Ferreira LCS, Shresta S
Cell reports 2020 Apr 28;31(4):107566

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Ovalbumin induces natural killer cells to secrete Th2 cytokines IL‑5 and IL‑13 in a mouse model of asthma.

Chen Z, Wang L
Molecular medicine reports 2019 Apr;19(4):3210-3216

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An Id2(RFP)-Reporter Mouse Redefines Innate Lymphoid Cell Precursor Potentials.

Xu W, Cherrier DE, Chea S, Vosshenrich C, Serafini N, Petit M, Liu P, Golub R, Di Santo JP
Immunity 2019 Apr 16;50(4):1054-1068.e3

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Dephosphorylated Polymerase I and Transcript Release Factor Prevents Allergic Asthma Exacerbations by Limiting IL-33 Release.

Ni Y, Hao J, Hou X, Du W, Yu Y, Chen T, Wei Z, Li Y, Zhu F, Wang S, Liang R, Li D, Lu Y, Liao K, Li B, Shi G
Frontiers in immunology 2018;9:1422

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Bcl11b is essential for licensing Th2 differentiation during helminth infection and allergic asthma.

Lorentsen KJ, Cho JJ, Luo X, Zuniga AN, Urban JF Jr, Zhou L, Gharaibeh R, Jobin C, Kladde MP, Avram D
Nature communications 2018 Apr 26;9(1):1679

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Toxoplasma Co-infection Prevents Th2 Differentiation and Leads to a Helminth-Specific Th1 Response.

Ahmed N, French T, Rausch S, Kühl A, Hemminger K, Dunay IR, Steinfelder S, Hartmann S
Frontiers in cellular and infection microbiology 2017;7:341

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Regulatory Innate Lymphoid Cells Control Innate Intestinal Inflammation.

Wang S, Xia P, Chen Y, Qu Y, Xiong Z, Ye B, Du Y, Tian Y, Yin Z, Xu Z, Fan Z
Cell 2017 Sep 21;171(1):201-216.e18

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Water-soluble chitosan inhibits nerve growth factor and attenuates allergic inflammation in mite allergen-induced allergic rhinitis.

Chen PC, Hsieh MH, Kuo WS, Kao HF, Hsu CL, Wang JY
The Journal of allergy and clinical immunology 2017 Oct;140(4):1146-1149.e8

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Intestinal helminth infection induces highly functional resident memory CD4(+) T cells in mice.

Steinfelder S, Rausch S, Michael D, Kühl AA, Hartmann S
European journal of immunology 2017 Feb;47(2):353-363

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Transcription Factors Oct-1 and GATA-3 Cooperatively Regulate Th2 Cytokine Gene Expression via the RHS5 within the Th2 Locus Control Region.

Kim K, Kim N, Lee GR
PloS one 2016;11(2):e0148576

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Characterization and Quantification of Innate Lymphoid Cell Subsets in Human Lung.

De Grove KC, Provoost S, Verhamme FM, Bracke KR, Joos GF, Maes T, Brusselle GG
PloS one 2016;11(1):e0145961

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Lack of autophagy induces steroid-resistant airway inflammation.

Suzuki Y, Maazi H, Sankaranarayanan I, Lam J, Khoo B, Soroosh P, Barbers RG, James Ou JH, Jung JU, Akbari O
The Journal of allergy and clinical immunology 2016 May;137(5):1382-1389.e9

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The IL-10 polarized cytokine pattern in innate and adaptive immunity cells contribute to the development of FVIII inhibitors.

Silveira AC, Santana MA, Ribeiro IG, Chaves DG, Martins-Filho OA
BMC hematology 2015;15(1):1

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TNF superfamily member TL1A elicits type 2 innate lymphoid cells at mucosal barriers.

Yu X, Pappu R, Ramirez-Carrozzi V, Ota N, Caplazi P, Zhang J, Yan D, Xu M, Lee WP, Grogan JL
Mucosal immunology 2014 May;7(3):730-40

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Thymic stromal lymphopoietin induces corticosteroid resistance in natural helper cells during airway inflammation.

Kabata H, Moro K, Fukunaga K, Suzuki Y, Miyata J, Masaki K, Betsuyaku T, Koyasu S, Asano K
Nature communications 2013;4:2675

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Group V secretory phospholipase A2 is involved in macrophage activation and is sufficient for macrophage effector functions in allergic pulmonary inflammation.

Ohta S, Imamura M, Xing W, Boyce JA, Balestrieri B
Journal of immunology (Baltimore, Md. : 1950) 2013 Jun 15;190(12):5927-38

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The acute environment, rather than T cell subset pre-commitment, regulates expression of the human T cell cytokine amphiregulin.

Qi Y, Operario DJ, Georas SN, Mosmann TR
PloS one 2012;7(6):e39072

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Lack of IL-4 receptor expression on T helper cells reduces T helper 2 cell polyfunctionality and confers resistance in allergic bronchopulmonary mycosis.

Müller U, Piehler D, Stenzel W, Köhler G, Frey O, Held J, Grahnert A, Richter T, Eschke M, Kamradt T, Brombacher F, Alber G
Mucosal immunology 2012 May;5(3):299-310

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Donor B cells in transplants augment clonal expansion and survival of pathogenic CD4+ T cells that mediate autoimmune-like chronic graft-versus-host disease.

Young JS, Wu T, Chen Y, Zhao D, Liu H, Yi T, Johnston H, Racine J, Li X, Wang A, Todorov I, Zeng D
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Transcription factor RORα is critical for nuocyte development.

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The characterization of four monoclonal antibodies specific for mouse IL-5 and development of mouse and human IL-5 enzyme-linked immunosorbent.

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Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were polarized under Th2 conditions and then unstimulated (left) or stimulated for 5 hours with Cell Stimulation Cocktail (Product # 00-4970-03) in the presence of Brefeldin A Solution (Product # 00-4506-51) (right). Cells were intracellularly stained with Anti-Human CD4 FITC (Product # 11-0048-42) and 0.125 µg of Anti-Human/Mouse IL-5 PE using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis.

Conjugate

Yellow dye

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 4 Frequencies of CD4 + T-cells expressing Gata3 and Th2 cytokines in asthma and helminth infection are reduced in Bcl11b fl/fl dLck-iCre mice by an intrinsic defect. a Flow cytometry analysis of IL-4 (left column), IL-5 (middle column), and IL-13 (right column) against GATA3 in the CD4 + T-cell population of the lung parenchyma of HDM asthma-induced mice. b Frequencies of IL-4 + ( n = 4), IL-5 + ( n = 4), and IL-13 + ( n = 8) (top panel) and GATA3 + ( n = 10) (bottom panel) CD4 + T-cells from the lungs of the indicated groups. Data are derived from two (IL-4, IL-5, and IL-13 data) or five (GATA3 data) independent experiments. c Flow cytometry analysis of IL-4 (left column), IL-5 (middle column), and IL-13 (right column) against GATA3 in the CD4 + T-cell population from the mesenteric (m)LNs of H. polygyrus infected mice. d Frequencies of IL-4 + ( n = 4), IL-5 + ( n = 4), and IL-13 + ( n = 4) (top panel) and GATA3 + ( n = 8) (bottom panel) CD4 + T-cells from mLNs of the indicated groups. Data are derived from two (IL-4, IL-5, and IL-13) or four (GATA3) independent experiments. e Flow cytometry analysis of GATA3 in CD4 + T-cells (left column) and IL-4 (middle and right columns) in the GATA3 hi (middle column) and GATA3 lo (right column) CD4 + T-cells from the mLNs of H. polygyrus bakeri infected mice. f The ratio of GATA3 hi /GATA3 lo CD4 + T-cells (top) and frequencies of IL-4 secreting cells within the GATA3 hi and GATA3 lo CD4 + T-cells (bottom) from the mLNs of H. poly

Conjugate

Yellow dye