Antibody data
- Antibody Data
- Antigen structure
- References [11]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [8]
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- Product number
- 46-7108-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IL-10 Monoclonal Antibody (JES3-9D7), PerCP-eFluor™ 710, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The JES3-9D7 monoclonal antibody reacts with human interleukin-10 (IL-10). Applications Reported: This JES3-9D7 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This JES3-9D7 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of stimulated normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Bufer Set (Product # 88-8824-00) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic)intracellular proteins. This can be used at 5 µL (0.03 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. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Rat
- Isotype
- IgG
- Antibody clone number
- JES3-9D7
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Early loss of T lymphocyte 4-1BB receptor expression is associated with higher short-term mortality in alcoholic hepatitis.
Cysticercus cellulosae Regulates T-Cell Responses and Interacts With the Host Immune System by Excreting and Secreting Antigens.
CAIX Regulates GBM Motility and TAM Adhesion and Polarization through EGFR/STAT3 under Hypoxic Conditions.
Multiple Sclerosis-Associated Changes in the Composition and Immune Functions of Spore-Forming Bacteria.
TIGIT expressing CD4+T cells represent a tumor-supportive T cell subset in chronic lymphocytic leukemia.
Heterologous Immunity between Adenoviruses and Hepatitis C Virus: A New Paradigm in HCV Immunity and Vaccines.
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Granzyme B production distinguishes recently activated CD8(+) memory cells from resting memory cells.
Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.
Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.
Role of the phosphatidylinositol 3 kinase-Akt pathway in the regulation of IL-10 and IL-12 by Porphyromonas gingivalis lipopolysaccharide.
Eriksen LL, Nielsen MA, Laursen TL, Deleuran B, Vilstrup H, Støy S
PloS one 2021;16(8):e0255574
PloS one 2021;16(8):e0255574
Cysticercus cellulosae Regulates T-Cell Responses and Interacts With the Host Immune System by Excreting and Secreting Antigens.
Fan X, Zhang Y, Ouyang R, Luo B, Li L, He W, Liu M, Jiang N, Yang F, Wang L, Zhou B
Frontiers in cellular and infection microbiology 2021;11:728222
Frontiers in cellular and infection microbiology 2021;11:728222
CAIX Regulates GBM Motility and TAM Adhesion and Polarization through EGFR/STAT3 under Hypoxic Conditions.
Huang BR, Liu YS, Lai SW, Lin HJ, Shen CK, Yang LY, Lu DY
International journal of molecular sciences 2020 Aug 14;21(16)
International journal of molecular sciences 2020 Aug 14;21(16)
Multiple Sclerosis-Associated Changes in the Composition and Immune Functions of Spore-Forming Bacteria.
Cekanaviciute E, Pröbstel AK, Thomann A, Runia TF, Casaccia P, Katz Sand I, Crabtree E, Singh S, Morrissey J, Barba P, Gomez R, Knight R, Mazmanian S, Graves J, Cree BAC, Zamvil SS, Baranzini SE
mSystems 2018 Nov-Dec;3(6)
mSystems 2018 Nov-Dec;3(6)
TIGIT expressing CD4+T cells represent a tumor-supportive T cell subset in chronic lymphocytic leukemia.
Catakovic K, Gassner FJ, Ratswohl C, Zaborsky N, Rebhandl S, Schubert M, Steiner M, Gutjahr JC, Pleyer L, Egle A, Hartmann TN, Greil R, Geisberger R
Oncoimmunology 2017;7(1):e1371399
Oncoimmunology 2017;7(1):e1371399
Heterologous Immunity between Adenoviruses and Hepatitis C Virus: A New Paradigm in HCV Immunity and Vaccines.
Singh S, Vedi S, Samrat SK, Li W, Kumar R, Agrawal B
PloS one 2016;11(1):e0146404
PloS one 2016;11(1):e0146404
Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.
Miles B, Miller SM, Folkvord JM, Kimball A, Chamanian M, Meditz AL, Arends T, McCarter MD, Levy DN, Rakasz EG, Skinner PJ, Connick E
Nature communications 2015 Oct 20;6:8608
Nature communications 2015 Oct 20;6:8608
Granzyme B production distinguishes recently activated CD8(+) memory cells from resting memory cells.
Nowacki TM, Kuerten S, Zhang W, Shive CL, Kreher CR, Boehm BO, Lehmann PV, Tary-Lehmann M
Cellular immunology 2007 May;247(1):36-48
Cellular immunology 2007 May;247(1):36-48
Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.
Sendide K, Deghmane AE, Pechkovsky D, Av-Gay Y, Talal A, Hmama Z
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32
Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.
Sendide K, Deghmane AE, Pechkovsky D, Av-Gay Y, Talal A, Hmama Z
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32
Role of the phosphatidylinositol 3 kinase-Akt pathway in the regulation of IL-10 and IL-12 by Porphyromonas gingivalis lipopolysaccharide.
Martin M, Schifferle RE, Cuesta N, Vogel SN, Katz J, Michalek SM
Journal of immunology (Baltimore, Md. : 1950) 2003 Jul 15;171(2):717-25
Journal of immunology (Baltimore, Md. : 1950) 2003 Jul 15;171(2):717-25
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood mononuclear cells were cultured for 6 days under Th2-polarizing conditions. The cells were unstimulated (left) or restimulated (right) with the Cell Stimulation Cocktail (plus protein transport inhibitors) (Product # 00-4975-03) then intracellularly stained with Anti-Human CD4 FITC (Product # 11-0049-42) and Anti-Human IL-10 PerCP-eFluor® 710 using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Total viable cells, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Fig 4 Cross-reactive CD4 + and CD8 + T cells obtained from Ad vector immunized mice produce cytokines upon ex vivo stimulation with various HCV proteins. Splenocytes obtained from Ad vector immunized mice were cultured with HCV core, NS3, NS4 or NS5 antigens at 5 mug/ml, and analyzed after 5 days for intracellular IFN-gamma and IL-10 expression profile of CD4 + and CD8 + T cells by flow cytometry. Data are obtained from a pool (n = 5) of spleen cells and are representative of two independent experiments.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig 5 Cross-reactive CD4 + and CD8 + T cells obtained from Ad vector immunized mice produce cytokines upon ex vivo stimulation with HCV peptides. Splenocytes obtained from Ad vector immunized mice were cultured with representative peptides derived from HCV core, NS3, NS4 or NS5 at 5 mug/ml each, and analyzed after 5 days for intracellular IFN-gamma and IL-10 expression profile of CD4 + and CD8 + T cells by flow cytometry. Data are obtained from a pool (n = 5) of spleen cells and are representative of two independent experiments.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 6 T FR exhibit an enhanced regulatory phenotype in ex vivo HIV infection. Tonsil cells were mock-, X4-, or R5-spinoculated and cultured under experimental conditions as indicated. T FR were then analysed for expression of regulatory receptors and cytokine production by intracellular cytokine staining. ( a ) Percentage of total (surface and intracellular) T FR CTLA-4 expression ( n =15). ( b ) Percentage of surface T FR LAG-3 expression ( n =8). ( c ) Production of IL-10 by T FR ( n =7). ( d ) Production of TGF-beta-1 (measured as LAP) by T FR ( n =5). The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman ( a , b ) or Mann-Whitney tests ( c , d ) and significance is denoted by asterisks where * P
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- FIG 2 Spore-forming bacteria from MS patients inhibit IL-10 + Treg differentiation in vitro . (A and B) Representative flow cytometry plots (A) and quantification (B) of CD4 + FoxP3 + Tregs within CD3 + lymphocytes differentiated in response to spore-forming bacteria isolated from controls or untreated MS patients. n = 7 PBMC donors; each dot represents an average response from PBMC donor to isolates from 6 control or MS bacterial donors. * * , P < 0.01, two-tailed repeated measures t test. (C and D) Representative flow cytometry plots (C) and quantification (D) of IL-10 + lymphocyte population within CD3 + CD4 + FoxP3 + Tregs differentiated in response to spore-forming bacteria isolated from controls or untreated MS patients. n = 6 bacterial donors per group. * , P < 0.05, two-tailed t test. Error bars, mean +- SEM. The experiment was repeated with nonoverlapping PBMC and bacterial donors and gave the same results. (E) Quantification of T effector cell proliferation in response to Tregs differentiated in the presence of spore-forming bacteria from MS patients or controls. n = 3 bacterial donors per group, each representing an average of 3 technical replicates. (F) Linear correlation between IL-10 + population within CD3 + CD4 + FoxP3 + Tregs and Clostridia-Bacilli relative abundances. R 2 = 0.214, P = 0.0459. Black dots, MS patients. Light gray dots, controls.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 4 The polarizations of monocyte and GBM progression in a co-cultured model under hypoxic conditions. ( A ) THP-1 monocytes were co-cultured with U87-green fluorescent protein (GFP) or U251-GFP under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of cluster of differentiation (CD) 206, arginase 1 (Arg1), interleukin (IL)-10, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha by flow cytometry. ( B ) The median fluorescence intensity (MFI) of CD206, Arg1, IL-10, IFN-gamma, and TNF-alpha in THP-1 monocytes in the co-cultured model under hypoxic conditions for 48 h. * p < 0.05 compared with the THP-1 monocyte group. Quantitative data are presented as the mean +- standard error (representative of n = 3). ( C ) U87-GFP and U251-GFP were treated with U104 (CAIX inhibitor) and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of CD206, Arg1, and IL-10 by flow cytometry. ( D ) U87-GFP or U251-GFP was treated with U104 and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The GFP-positive gated GBM was analyzed to assess the levels of programmed death ligand 1 (PD-L1) by flow cytometry. ( E ) The levels of CAIX, PD-L1, tumor growth factor-beta, and c
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- Invitrogen Antibodies (provider)
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- 10.1371/journal.pone.0255574.g002 Fig 2 Cytokine production in 4-1BB + and 4-1BB - CD4 + T lymphocytes. Flow cytometry of peripheral blood mononuclear cells stimulated for 48 hours with anti-CD3 and anti-CD28 given PMA and brefaldin-A for the last 4 hours. (A) Typical flow cytometry plot of 4-1BB + CD4 + T lymphocytes from a patient with alcoholic hepatitis showing interferon-gamma (IFNy) on x-axis and interleukin-10 (IL-10) on y-axis. Frequencies of interferon-gamma + (B, n = 6) and IL-10 + (C, n = 3)) 4-1BB + and 4-1BB - CD4 + T lymphocytes in patients with alcoholic hepatitis. Differences between groups compared using paried T test. Graphs shown as median with interquartile range.
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- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 5 IL-10 + lymphocyte assay. The production of IL-10 + lymphocyte subsets by flow cytometry. (A) Both Cysticercus cellulosae ESA and CA induced the production of IL-10 + lymphocyte. (B) CD4 + T lymphocyte was the main source Cysticercus cellulosae ESA and CA-induced IL-10 expression. (C) In the presence or absence of LPS, Cysticercus cellulosae ESA and CA significantly inhibited the expression frequency of CD8 + IL-10 + lymphocyte. (D) Cysticercus cellulosae ESA could be induced the expression of CD4 - CD8 - IL-10 + cell under the presence or absence of LPS. While, CA only decreased the expression in the presence of LPS. (E) Similar to (C) , the production of IL-10 in CD4 + CD8 + lymphocyte was inhibited by Cysticercus cellulosae ESA. All dates were shown as the means +- SD, * P < 0.05, ** P < 0.01, *** P < 0.001.
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- Invitrogen Antibodies (provider)
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- Figure 6 The production of Foxp3 + IL-10 + lymphocyte. The co-expressing Foxp3 and IL-10 cell were detected via tricolor flow cytometry, which shown that the production of Foxp3 + IL-10 + lymphocyte was inhibited by Cysticercus cellulosae ESA. Besides, CA had significantly decreased the production only under the presence of LPS. Results were expressed as the mean+- SD, * P < 0.05.