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Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Mouse
Cited:
Human, Mouse, Rat, Transgenic Mouse
Applications
Validated:
Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Bioassay, ELISA Capture, ELISA Development, ELISA Development (Capture), ELISpot Development, Functional Assay, Luminex Development, Tissue Culture
Label
Unconjugated
Antibody Source
Monoclonal Rat IgG1 Clone # JES052A5
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Product Specifications
Immunogen
E. coli-derived recombinant mouse IL-10
Accession # P18893
Accession # P18893
Specificity
Detects mouse IL-10 in sandwich ELISAs and Western blots. In sandwich ELISAs, less than 8% cross-reactivity with recombinant rat IL-10 and 1% cross-reactivity with recombinant cotton rat IL‑10 is observed. In sandwich ELISAs, no cross-reactivity with recombinant human (rh) IL-10, recombinant mouse IL-10 R, rhIL‑10 R, recombinant canine IL-10, recombinant feline IL-10, or recombinant porcine IL-10 is observed.
Clonality
Monoclonal
Host
Rat
Isotype
IgG1
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Mouse IL-10 Antibody (JES052A5)
Detection of Recombinant Mouse IL‑10 by Western Blot.
Western blot shows 25 ng of Recombinant Mouse IL-10 (Catalog # 417-ML), Recombinant Human IL-10 (Catalog # 217-IL), Recombinant Rat IL-10 (Catalog # 522-RLB), and Recombinant Human IL-26/AK155 Monomer (Catalog # 1375-IL). PVDF Membrane was probed with 1 µg/mL of Rat Anti-Mouse IL-10 Monoclonal Antibody (Catalog # MAB417) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for IL-10 at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Cell Proliferation Induced by IL‑10 and Neutralization by Mouse IL‑10 Antibody.
Recombinant Mouse IL-10 (Catalog # 417-ML) stimulates proliferation in the MC/9-2 mouse mast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Mouse IL-10 (2.5 ng/mL) is neutralized (green line) by increasing concen-trations of Rat Anti-Mouse IL-10 Monoclonal Antibody (Catalog # MAB417). The ND50 is typically 0.005-0.015 µg/mL.
Detection of Mouse IL-10 by Western Blot
Imipramine downregulates an M2-like program in TAMs(A) Western blot analysis of ARG1 and IL-10 in single tumors treated or not with IM or anti-VEGF.(B) Flow cytometry analysis of ARG1 and IL-10 expression in GBM tumors treated for 1 week. Ctrl (n = 9 tumors), IM (n = 6), anti-VEGF (n = 6), anti-VEGFR2 (n = 6), Axitinib (n = 6). Macrophages were gated as CD45+CD11b+Ly6C-Ly6G−.(C) Expression of ARG1 and IL-10 in microglia (CD49d−) and MDMs (CD49d+) assessed by FACS in untreated tumors (n = 5).(D) Expression of MHC-II within microglia as assessed by flow cytometry. Ctrl (n = 4 tumors) and IM + anti-VEGF (n = 4).(E) Ex vivo co-cultures of tumoral CD11b cells and activated splenic CFSE-labeled CD8 or CD4 T cells. Each dot represents the average of two or three technical replicates. T cells alone (n = 4), Ctrl co-culture (n = 5), anti-VEGF (n = 3), IM (n = 4), IM + anti-VEGF (n = 4).(F) Analysis of the M2-like program in cytokine-polarized macrophages as assessed by qRT-PCR analysis of Ctrl and IM-treated M2-like BMDMs. Expression is normalized to 18S statistics by Welch’s t test. Each dot represents an individual sample. Data are representative of three independent experiments.(G) Analysis of the M1-like program in BMDMs assessed by FACS. Each dot represents an individual replicate.(H) Expression of Hrh1 mRNA normalized to 18S in ex vivo M1-and M2-polarized BMDMs, either untreated or IM treated for 24 h. Each dot represents an individual sample. Data are representative of three independent experiments.(I) mRNA expression of Hrh1 in FACS-sorted microglia or MDMs from Ctrl and IM-treated tumors.(J) mRNA expression of Arg1, Chil3, and Il10 in M2-polarized macrophages that were transfected with siCtrl or two different siHrh1 constructs. Cells were treated with 40 μm IM for 24 h. Data are representative of two independent experiments.(K) Western blot analysis of MRC1 and ARG1 expression of siRNA-transfected M2 BMDMs. Data are representative of two independent experiments.(L)
Detection of Mouse IL-10 by Immunocytochemistry/Immunofluorescence
Lymphocyte infiltrate in tumors from IL-10 deficient mice (A, C and D) or from anti-IL10/IL-10R neutralizing antibody treated mice (B, E and F). A. Single cell suspensions were stained with the indicated antibodies plus anti-CD45 and anti-F4/80. Lymphocytes were analyzed in the CD45+F4/80- population. B. After 6 hours incubation in tissue culture flasks, non adherent cells from total tumor suspension were harvested and stained with the indicated antibodies, tetramer and anti-CD45. C and E. Immunofluorescence of cryo-sections from tumors harvested from IL-10 KO and wild type mice or neutralizing anti-IL10/IL-1R and irrelevant IgG antibodies, as indicated. Sections were stained with anti-CD8 (in FITC, green) and PE (red) conjugated HPV16 E749-57 tetramer - E749-57 Tet, and irrelevant tetramer - Irrelevant Tet. Cells were counterstained with DAPI (blue). Arrowheads indicate double positive cells in yellow, arrows indicate cells positive only for CD8. D and F. Granzyme B expression in tumor cryo-sections from IL-10 KO and wild type mice or mice treated with anti-IL10/IL-10R neutralizing antibodies and irrelevant IgG, as indicated. Sections were stained with anti-CD8 (green), followed by fixation, permeabilization and staining with anti-Granzyme B (red). Tissue was counterstained with DAPI. Solid arrows indicate double positive cells, arrowhead indicate CD8 only positive cell. Insets show details of cells positive only for CD8 (green) or double positive CD8/tetramer (C and E), CD8/Granzyme B (D and F) in yellow or orange due to the sobreposition of emission from FITC and PE (tetramer) or alexa 594 (Granzyme B). Image collected and cropped by CiteAb from the following publication (https://bmcimmunol.biomedcentral.com/articles/10.1186/1471-2172-11-27), licensed under a CC-BY license. Not internally tested by R&D Systems.
Mouse IL-10 ELISA Standard Curve
Recombinant Mouse IL‑10 (Catalog # 417-ML) was serially diluted and captured by Rat Anti-Mouse IL‑10 Monoclonal Antibody (Catalog # MAB417) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Mouse IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-417-NA) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)Applications for Mouse IL-10 Antibody (JES052A5)
Application
Recommended Usage
Western Blot
1 µg/mL
Sample: Recombinant Mouse IL‑10 (Catalog # 417-ML)
Sample: Recombinant Mouse IL‑10 (Catalog # 417-ML)
Neutralization
Measured by its ability to neutralize IL‑10-induced proliferation in the MC/9‑2 mouse mast cell line. Thompson-Snipes, L. et al. (1991) J. Exp. Med. 173:507. The Neutralization Dose (ND50) is typically 0.005-0.015 µg/mL in the presence of 2.5 ng/mL Recombinant Mouse IL‑10.
Mouse IL-10 Sandwich Immunoassay
Please Note: Optimal dilutions of this antibody should be experimentally determined.
Reviewed Applications
Read 2 reviews rated 5 using MAB417 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: IL-10
Long Name
Interleukin 10
Alternate Names
CSIF, GVHDS, IL10, IL10A, TGIF
Entrez Gene IDs
Gene Symbol
IL10
UniProt
Additional IL-10 Products
Product Documents for Mouse IL-10 Antibody (JES052A5)
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Mouse IL-10 Antibody (JES052A5)
For research use only
Related Research Areas
Citations for Mouse IL-10 Antibody (JES052A5)
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Customer Reviews for Mouse IL-10 Antibody (JES052A5) (2)
5 out of 5
2 Customer Ratings
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Application: Flow CytometrySample Tested: T cellsSpecies: MouseVerified Customer | Posted 03/16/2022
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Application: Western BlotSample Tested: Peritoneal macrophagesSpecies: MouseVerified Customer | Posted 11/13/2020
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
