Clone:
JES3-9D7
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
rat IgG1κ, rat IgG1
Applications:
ICFC, FA
Alternative names:
IL10, CSIF, GVHDS, TGIF, IL10A

Extended validation for IL-10 Antibody, anti-human

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with JES3-9D7
2050B-
JES3-12G8-
JES3-19F1++
REA842++
Cells were incubated with an excess of purified unconjugated IL-10 (JES3-9D7) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for IL-10. Human peripheral blood mononuclear cells (PBMCs) were stimulated with CytoStim for 1 hour, followed by an incubation with 1µg/mL brefeldin A for 5 hours. Cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized followed by a staining with IL-10 antibodies. As a control, IL-10 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for IL-10 Antibody, anti-human

Overview

Interleukin 10 (IL‑10) is a cytokine predominantly secreted by CD4
+
memory and effector T cells and antigen-presenting cells, for example, monocytes/macrophages and dendritic cells. IL-10 has important suppressive functions on immune responses and is believed to be involved in the maintenance of tolerance. IL-10 blocks activation of cytokine synthesis by Tʜ1 cells, activated monocytes, and NK cells. It can stimulate immunoglobulin production by B cells.
The Anti-IL‑10 antibodies has been designed for intracellular staining of IL‑10–producing cells. Cells can be stimulated for IL‑10 production, for example, by polyclonal stimulation with mitogens. For induction of IL‑10 production by antigen-specific T cells, cells are re-stimulated with the respective antigen. IL‑10 can be accumulated in the cells by addition of secretion inhibitors like brefeldin A. After fixation and permeabilization of the cell sample, IL‑10–producing cells can be stained intracellularly with Anti-IL‑10 antibodies. Staining of surface markers allows simultaneous flow cytometric analysis of subsets and activation status of the IL‑10–producing cells.
Magnetically enriched cells can be stained intracellularly for IL‑10 production directly on the MACS
®
Column. This procedure ensures higher sensitivity of detection and minimizes loss of cells during washing procedures. The protocol is useful for cytokine analysis of rare cells, for example, CD4
+
T cells in HIV patients, or other cell sources than peripheral blood mononuclear cells (PBMCs), e.g., bronchoalveolar lavages or synovial fluids.

Alternative names

IL10, CSIF, GVHDS, TGIF, IL10A

Detailed product information

Technical specifications

CloneJES3-9D7
Clonalitymonoclonal
Isotyperat IgG1κ, rat IgG1
Isotype controlIsotype Control Antibody, rat IgG1
Hostrat
Type of antibodyPrimary antibodies, Functional-grade antibodies
Specieshuman, non-human primate
Cross-reactivity
rhesus monkey (
Macaca mulatta
)
AntigenIL-10
Alternative names of antigenIL10, CSIF, GVHDS, TGIF, IL10A
Molecular mass of antigen [kDa]19
Entrez Gene ID3586
RRIDAB_2857541, AB_2811315, AB_2811312, AB_2660623, AB_2660631, AB_2660632, AB_2857542

Resources for IL-10 Antibody, anti-human

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

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