14-7108-81

antibody from Invitrogen Antibodies

Targeting: IL10 CSIF, IL-10, IL10A, TGIF

Provider product page for 14-7108-81

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Antibody data

Antibody Data
Antigen structure
References [34]
Comments [0]
Validations
Western blot [1]
Other assay [12]

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Product number: 14-7108-81 - Provider product page
Provider: Invitrogen Antibodies
Product name: IL-10 Monoclonal Antibody (JES3-9D7), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The JES3-9D7 monoclonal antibody reacts with human interleukin-10 (IL-10). Applications Reported: The JES3-9D7 antibody has been reported for use in capture of human IL-10 by ELISA, intracellular flow cytometric analysis, and for neutralizing IL-10 bioactivity. (Use Functional Grade purified for functional studies Product # 16-7108). Applications Tested: The JES3-9D7 antibody has been tested as the capture antibody in a sandwich ELISA for analysis of human Interleukin-10 (IL-10) in combination with the biotin JES3-12G8 (Product # 13-7109-81) antibody for detection and recombinant human IL-10 (14-8109) as the standard. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/mL. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 1000 pg/mL - 8 pg/mL should be included in each ELISA plate. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Rat
Isotype: IgG
Antibody clone number: JES3-9D7
Vial size: 50 µg
Concentration: 0.5 mg/mL
Storage: 4° C

IL10 protein structure - 14-7108-81 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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

Supportive validation

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Experimental details: Figure 5 AhR-positive MCs express the tolerogenic IL-10 cytokine in ETs. ( A ) Immunofluorescence triple staining showing that several tryptase-expressing MCs (Tryp, green) co-express both AhR (white) and IL-10 (red) (arrows). Scale bar, 25 um. ( B ) A higher magnification view of the boxed area in (A) comparing uncoupled fluorescence micrographs of three MCs. ( C ) The percentage of triple-positive cells among tryptase-expressing MCs in 10 individual control and ET samples is shown. Cells were detected and counted from ten x400 high-power microscopic fields in each sample. Endometrium median 33% (IQR 31-37%); ovarian median 55% (IQR 53-73%). * p

Supportive validation

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Experimental details: Figure 6 (A) Ovarian ET cytokine expression upon treatment with an AhR antagonist. IL-4, IL-10, IL-12, IL-17, GM-CSF, IFN- , MCP-1/CCL2, MIP-1 /CCL4 and TNF-alpha protein expression in supernatants of untreated (control) or ANF-treated ovarian ET cultures measured by ELISA 48 hr after exposure to the AhR-antagonist ANF (15 uM). Control protein concentration was assigned a value of 1 and is illustrated as a horizontal line for presentation purposes. Sample size was 4 patients/group. The values are the mean and 95% confidence intervals. * p

Supportive validation

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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.

Supportive validation

Submitted by: 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

Supportive validation

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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Characterization and immunstimulation of Ad-hCD40L-transduced DC. a Map of Ad-hCD40L. LITR left-inverted terminal repeat, ES encapsidation signal, CMV cytomegalovirus immediate early promoter, MCS multiple cloning site, PolyA polyadenylation signal, Ad5del.E1/E3 human adenovirus type 5 sequence with deletion of E1/E3 genes, RITR right-inverted terminal repeat. b Light (left) and fluorescence (right) microscopy of DC 48 h after transduction with Ad-GFP (magnification x 10). c Light microscopy of DC 48 h after transduction with Ad-Mock (left) or Ad-hCD40L (right). Ad-hCD40L-transduced DC form cell aggregates (magnification x 10). d Cytokine amounts in the supernatant of DC 48 h after adenoviral transduction with Ad-hCD40L, Ad-Mock or non-transduced measured by ELISA. (* p < 0.05), n.d. = non-detectable. Data represent means +- SEM of six different experiments. e Expression of sCD40L 48 h after transduction of DC with Ad-hCD40L with different MOIs, ( n = 4-5). Data represent means +- SEM of three different experiments

Supportive validation

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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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.