Invitrogen

IL-10 Monoclonal Antibody (JES5-16E3), Biotin, Functional Grade, eBioscience™

Product Image Gallery

IL-10 Antibody in Flow Cytometry (Flow) — Figure 7. ERK2 phosphorylation of RORgammat protects against exacerbated DSS-induced colitis (A) HEK293 cells transfected with empty or 2xFLAG-RORgammat expression constructs for 16 h were treated with dimethylsulfoxide (DMSO, Veh) or ERK inhibitor (PD0325901, 5muM) and stimulated with the indicated cytokines. RORgammat was captured by anti-FLAG beads and pS182 status was detected by immunoblot. (B) HEK293 cells transfected with empty or 2xFLAG-RORgammat expression constructs for 16 h. RORgammat was captured by anti-FLAG beads and incubated with recombinant ERK2 in vitro . Phospho-S182 status was detected by immunoblot. This experiment was repeated twice with similar results. (C) Representative images from proximity ligation assay (PLA) indicating interactions between RORgammat and ERK1/2 in wild-type cultured Th17 cells. (D) Representative flow cytometry analysis of IL-17A expression in wild-type Th17 cells polarized under IL-6 (20 ng/mL), IL-1beta (20 ng/mL), and IL-23 (25 ng/mL) in the presence or absence of ERK inhibitor (PD0325901, 5muM) at 48 h. Cells were harvested and analyzed at 72 h. (E) Representative flow cytometry analysis of IL-10 expression in wild-type RORgammat + Treg cells polarized under IL-6 (20 ng/mL), IL-1beta (20 ng/mL), and TGF-beta (5 ng/mL) in the presence or absence of ERK inhibitor (PD0325901, 5muM) at 48 h. Cells were harvested and analyzed at 72 h. (F) Weight changes in RORgammat WT (left) and RORgammat S182A (right) cohoused littermates challeng

IL-10 Antibody in Flow Cytometry, ELISA (Flow, ELISA) — Figure 4 Co-culture of A20 B lymphoma cells and mesenchymal stem cells (MSCs) causes a decrease in the levels of interleukin (IL)-10 in the supernatant. A20 cells (1x10 5 ) were cultured in 6-well plates in the presence or absence of MSCs at ratio 1:1 for 24, 48, or 72 h. (A) IL-10 levels in A20 culture supernatants were measured by ELISA in the presence or absence of MSCs for 72 h. (B) The fraction of CD19-positive A20 cells expressing intracellular IL-10 was measured by flow cytometry. (C) The levels of transforming growth factor (TGF)-beta, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma in the culture supernatants were measured by ELISA. (D) The fraction of cells containing intracellular IL-10 was measured by flow cytometry. Results are expressed as the means +- SD of 3 independent experiments. * P<0.05 indicates statistical significance.

IL-10 Antibody in Neutralization (Neu) — Figure 6 TGF-beta1 secreted from Tregs induces Il-17rb expression of breast cancer cells via Smad2/3/4 signaling A CD4 + CD25 + Tregs were isolated from inguinal LN of BALB/c mice injected with 4T1 cells for 3 weeks and were transwell co-cultured with 4T1 cells in the presence of control IgG or neutralizing antibodies against IL-10 or TGF-beta1. After 5-day co-culture, 4T1 cells at lower well were analyzed for Il-17rb expression by RT-qPCR analysis. Gapdh was used as internal control. B, C Western blotting analyzed Il-17rb in 4T1 cells treated with recombinant TGF-beta1 (B) or IL-10 (C) proteins for 5 days. D Western blotting analyzed Il-17rb in shLacZ or shTgfbr1 4T1 cells treated with recombinant TGF-beta1 for 5 days. E 4T1-injected BALB/c mice were sacrificed at 3 weeks postinitial injection. Total cells isolated from inguinal lymph node tissues were transwell co-cultured with 4T1 cells. After 5-day co-culture, soft-agar colony-forming activity was determined in co-cultured shLacZ or shTgfbr1 4T1 cells at lower well. (5 x 10 2 cells/well, n = 6 wells per group). F-H Western blotting analyzed Il-17rb in shLacZ, shSmad2 (F), shSmad3 (G), or shSmad4 (H) 4T1 cells treated with recombinant TGF-beta1 for 5 days. I, J WT 4T1 (I) or Il-17rb Del 4T1 (J) cells were treated with or without recombinant TGF-beta1 for 5 days. After treatment, nuclear translocation of NF-kappaB p65 was assayed by Western blotting in cells treated with recombinant mouse IL-17B for 2 h. Nuclear Matrix

Product Details

36-7101-85

Applications	Tested Dilution	Publications
Flow Cytometry (Flow)	-	View 16 publications 16 publications
ELISA (ELISA)	-	View 2 publications 2 publications
Neutralization (Neu)	-	View 1 publication 1 publication
Functional Assay (FN)	Assay-Dependent	-

Product Specifications
Species Reactivity	Mouse
Published species	Human, Mouse
Host/Isotype	Rat / IgG2b, kappa
Class	Monoclonal
Type	Antibody
Clone	JES5-16E3
Conjugate	Biotin , Functional Grade
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2
Contains	no preservative
Storage conditions	4° C
RRID	AB_469762

Product Specific Information

Description: The JES5-16E3 antibody reacts with mouse interleukin-10 (IL-10). Mouse IL-10 is an ~18 kDa factor also known as Cytokine Synthesis Inhibitory Factor (CSIF). In the mouse, Th2 cells, B1 cells, macrophages, and keratinocytes are the major cell subsets that produce IL-10. IL-10 inhibits synthesis of Th1 cytokines and proliferation of T cells, and acts as a costimulatory signal for mast cells, developing thymocytes and the Th2 response.

Applications Reported: The Functional Grade Biotin anti-mouse IL-10 antibody (clone JES5-16E3) has been reported for use as the in vivo capture antibody for the In Vivo Cytokine Capture Assay (IVCCA). The IVCCA facilitates measurement of cytokines in serum by increasing their in vivo half lives dramatically. This increases the sensitivity of measurement of in vivo cytokine production 30- to 1000- fold.

For detailed methodology in this application, please refer to Finkelman, F., S. Morris, T. Orekhova, and D. Sehy. 2003. The In Vivo Cytokine Capture Assay for measurement of cytokine production in the mouse. In Current Protocols in Immunology. Unit 6.28. J. Coligan, A. Kruisbeek, D. Margulies, E. Shevach, and W. Strober, eds. John Wiley and Sons, New York.

Applications Tested: The Functional Grade Biotin JES5-16E3 antibody has been tested by LAL assay to ensure it is free of endotoxin, and has been tested as the detection antibody in a sandwich ELISA for analysis of mouse IL-10 in combination with the purified JES5-2A5 (14-7102) antibody for capture and recombinant mouse IL-10 (14-8101) as the standard.A suitable range of concentrations of the FG Biotin JES5-16E3 antibody for ELISA detection is 0.5-2 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 4000 pg/mL - 30 pg/mL should be included in each ELISA plate.

Endotoxin: Less than 0.05 ng/µg antibody as determined by the LAL assay.

Storage and handling: Use in a sterile environment.

Target Information

Interleukin 10 (IL-10, CSIF) is an anti-inflammatory cytokine mainly produced by macrophages and Th2 cells. IL-10 is an 18.6 kDa protein of 160 amino acid residues that shares over 80% sequence homology with the Epstein-Barr Virus protein (BCRFI). The reported biological activities of IL-10, which maybe interrelated, include inhibition of macrophage-mediated cytokine synthesis, suppression of the delayed-type hyper-sensitivity response, and stimulation of the Th2 cell response which results in elevated antibody production. IL-10 functions by inhibiting pro-inflammatory cytokines made by macrophages and regulatory T cells including IFN-gamma, TNF-alpha, IL-2, and IL-3, IL-4, and GM-CSF. IL-10 is also known to suppress antigen presentation on antigen presenting cells, enhances B cell survival, proliferation, and antibody production. IL-10 can block NF-kappa B activity, and is involved in the regulation of the JAK-STAT signaling pathway. While human IL-10 can act on murine cells, murine IL-10 cannot act on human cells. Binds to a retinoid X receptor (RXR) responsive element from the cellular retinol-binding protein II promoter (CRBPII-RXRE). IL-10 inhibits the 9-cis-retinoic acid-dependent RXR alpha transcription activation of the retinoic acid responsive element. Knockout studies in mice suggest the function of this cytokine as an essential immunoregulator in the intestinal tract.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

RORγt phosphorylation protects against T cell-mediated inflammation.

Cell reports

Ma S,Patel SA,Abe Y,Chen N,Patel PR,Cho BS,Abbasi N,Zeng S,Schnabl B,Chang JT,Huang WJM

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Tue Mar 15 00:00:00 EDT 2022

Milk-derived extracellular vesicles alleviate ulcerative colitis by regulating the gut immunity and reshaping the gut microbiota.

Theranostics

Tong L,Hao H,Zhang Z,Lv Y,Liang X,Liu Q,Liu T,Gong P,Zhang L,Cao F,Pastorin G,Lee CN,Chen X,Wang JW,Yi H

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Fri Jan 28 00:00:00 EST 2022

Characterisation of Macrophage Polarisation in Mice Infected with Ninoa Strain of Trypanosoma cruzi.

Pathogens (Basel, Switzerland)

Medina-Buelvas DM,Rodríguez-Sosa M,Vega L

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Sat Nov 06 00:00:00 EDT 2021

Gut epithelial IL-27 confers intestinal immunity through the induction of intraepithelial lymphocytes.

The Journal of experimental medicine

Lin CH,Chen MC,Lin LL,Christian DA,Min B,Hunter CA,Lu LF

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Mon Nov 01 00:00:00 EDT 2021

Irradiated whole-cell vaccine suppresses hepatocellular carcinoma growth in mice via Th9 cells.

Oncology letters

Chen J,Ding Y,Huang F,Lan R,Wang Z,Huang W,Chen R,Wu B,Fu L,Yang Y,Liu J,Hong J,Zhang W,Zhang L

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Sat May 01 00:00:00 EDT 2021

TGF-β1 secreted by Tregs in lymph nodes promotes breast cancer malignancy via up-regulation of IL-17RB.

EMBO molecular medicine

Huang SC,Wei PC,Hwang-Verslues WW,Kuo WH,Jeng YM,Hu CM,Shew JY,Huang CS,Chang KJ,Lee EY,Lee WH

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Neutralization

Fri Dec 01 00:00:00 EST 2017

Excessive expression of miR-27 impairs Treg-mediated immunological tolerance.

The Journal of clinical investigation

Cruz LO,Hashemifar SS,Wu CJ,Cho S,Nguyen DT,Lin LL,Khan AA,Lu LF

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Wed Feb 01 00:00:00 EST 2017

Heterologous Immunity between Adenoviruses and Hepatitis C Virus: A New Paradigm in HCV Immunity and Vaccines.

PloS one

Singh S,Vedi S,Samrat SK,Li W,Kumar R,Agrawal B

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Tue Jul 19 00:00:00 EDT 2016

NFATc1 supports imiquimod-induced skin inflammation by suppressing IL-10 synthesis in B cells.

Nature communications

Alrefai H,Muhammad K,Rudolf R,Pham DA,Klein-Hessling S,Patra AK,Avots A,Bukur V,Sahin U,Tenzer S,Goebeler M,Kerstan A,Serfling E

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Wed May 25 00:00:00 EDT 2016

Maternal-Derived Hepatitis B Virus e Antigen Alters Macrophage Function in Offspring to Drive Viral Persistence after Vertical Transmission.

Immunity

Tian Y,Kuo CF,Akbari O,Ou JH

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Tue May 17 00:00:00 EDT 2016

Towards programming immune tolerance through geometric manipulation of phosphatidylserine.

Biomaterials

Roberts RA,Eitas TK,Byrne JD,Johnson BM,Short PJ,McKinnon KP,Reisdorf S,Luft JC,DeSimone JM,Ting JP

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Tue Dec 01 00:00:00 EST 2015

Tristetraprolin Limits Inflammatory Cytokine Production in Tumor-Associated Macrophages in an mRNA Decay-Independent Manner.

Cancer research

Kratochvill F,Gratz N,Qualls JE,Van De Velde LA,Chi H,Kovarik P,Murray PJ

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Sat Aug 01 00:00:00 EDT 2015

Mouse bone marrow-derived mesenchymal stem cells inhibit leukemia/lymphoma cell proliferation in vitro and in a mouse model of allogeneic bone marrow transplant.

International journal of molecular medicine

Song N,Gao L,Qiu H,Huang C,Cheng H,Zhou H,Lv S,Chen L,Wang J

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Wed Jul 01 00:00:00 EDT 2015

Mouse bone marrow-derived mesenchymal stem cells inhibit leukemia/lymphoma cell proliferation in vitro and in a mouse model of allogeneic bone marrow transplant.

International journal of molecular medicine

Song N,Gao L,Qiu H,Huang C,Cheng H,Zhou H,Lv S,Chen L,Wang J

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Wed Jul 01 00:00:00 EDT 2015

Nizatidine, a small molecular compound, enhances killed H5N1 vaccine cell-mediated responses and protects mice from lethal viral challenge.

Human vaccines & immunotherapeutics

Wang S,Wu B,Xue J,Wang M,Chen R,Wang B

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Mon Jan 05 00:00:00 EST 2015

BCG vaccine mediated reduction in the MHC-II expression of macrophages and dendritic cells is reversed by activation of Toll-like receptors 7 and 9.

Cellular immunology

Bakhru P,Sirisaengtaksin N,Soudani E,Mukherjee S,Khan A,Jagannath C

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Wed Jan 01 00:00:00 EST 2014

Genetic deletion of chemokine receptor Ccr7 exacerbates atherogenesis in ApoE-deficient mice.

Cardiovascular research

Wan W,Lionakis MS,Liu Q,Roffê E,Murphy PM

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Fri Mar 01 00:00:00 EST 2013

CX(3)CR1(+) macrophages support IL-22 production by innate lymphoid cells during infection with Citrobacter rodentium.

Mucosal immunology

Manta C,Heupel E,Radulovic K,Rossini V,Garbi N,Riedel CU,Niess JH

Published figure using IL-10 monoclonal antibody (Product # 36-7101-85) in Flow Cytometry

Tue Jan 01 00:00:00 EST 2013

Requirements for growth and IL-10 expression of highly purified human T regulatory cells.

Journal of clinical immunology

Bonacci B,Edwards B,Jia S,Williams CB,Hessner MJ,Gauld SB,Verbsky JW

36-7101-85 was used in Enzyme-linked immunosorbent assay to examine the requirements for growth and cytokine expression from highly purified human regulatory T cells, and study the signal transduction events of these cells.

Mon Oct 01 00:00:00 EDT 2012

Bioinformatics

Protein Aliases: CSIF; Cytokine synthesis inhibitory factor; H-IL-10; il 10; IL-10; ILN; Interleukin; Interleukin-10; Interleukin10; MGC126450; MGC126451; RP11-262N9.1

Gene Aliases: CSIF; Il-10; Il10

UniProt ID: (Mouse) P18893

Entrez Gene ID: (Mouse) 16153

Function(s)