Key Product Details

Species Reactivity

Validated:

Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Fluorescence Microscopy, In vivo assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
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Product Specifications

Immunogen

E. coli-derived recombinant rat IL‑10
Ser19-Asn178
Accession # P29456

Specificity

Detects mouse, rat, and cotton rat IL-10 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human IL-10, recombinant guinea pig IL-10, recombinant viral IL-10 and recombinant canine IL-10 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Mouse/Rat IL‑10 Antibody

Detection of Recombinant Mouse and Rat IL-10 antibody by Western Blot.

Detection of Recombinant Mouse and Rat 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 0.1 µg/mL of Goat Anti-Mouse/Rat IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF519) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for IL-10 at approximately 16 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 Rat IL‑10 Antibody.

Cell Proliferation Induced by IL‑10 and Neutralization by Rat IL‑10 Antibody.

Recombinant Rat IL-10 (Catalog # 522-RL) stimulates proliferation in the MC/9-2 mouse mast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL-10 (40 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF519). The ND50 is typically 0.3-1.0 µg/mL.

IL‑10 in Mouse Splenocytes.

IL‑10 was detected in immersion fixed mouse splenocytes using Goat Anti-Mouse/Rat IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF519) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Mouse/Rat IL‑10 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse IL‑10 (Catalog # 417-ML) and Recombinant Rat IL‑10 (Catalog # 522-RLB)

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 0.3-1.0 µg/mL in the presence of 40 ng/mL Recombinant Rat IL‑10.

Reviewed Applications

Read 5 reviews rated 4.6 using AF519 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IL-10

Interleukin 10, also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 family of alpha -helical cytokines that also includes IL-19, IL‑20, IL‑22, and IL-24 (1, 2). IL-10 is secreted by many activated hematopoietic cell types as well as hepatic stellate cells, keratinocytes, and placental cytotrophoblasts (2‑5). Mature rat IL‑10 shares 85% amino acid sequence identity with mouse and 71%‑79% amino acid sequence identity with bovine, canine, equine, feline, human, ovine, and porcine IL-10. Whereas human IL-10 is active on mouse cells, mouse IL-10 does not act on human cells (6, 7). IL-10 is a 178 amino acid molecule that contains two intrachain disulfide bridges and is expressed as a 36 kDa noncovalently associated homodimer (8‑10). The IL-10 dimer binds to two IL‑10 R alpha /IL‑10 R1 chains, resulting in recruitment of two IL-10 R beta /IL-10R2 chains and activation of a signaling cascade involving JAK1, TYK2, and STAT3 (11). IL‑10 R beta does not bind IL-10 by itself but is required for signal transduction (1). IL-10 R beta also associates with IL‑20 R alpha, IL-22 R alpha, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL-29 (12‑14). IL-10 is a critical molecule in the control of viral infections and allergic and autoimmune inflammation (15‑17). It promotes phagocytic uptake and Th2 responses but suppresses antigen presentation and Th1 proinflammatory responses (2).

References

  1. Pestka, S. et al. (2004) Annu. Rev. Immunol. 22:929.
  2. O’Garra, A. and P. Vieira (2007) Nat. Rev. Immunol. 7:425.
  3. Mathurin, P. et al. (2002) Am. J. Physiol. Gastrointest. Liver Physiol. 282:G981.
  4. Grewe, M. et al. (1995) J. Invest. Dermatol. 104:3.
  5. Szony, B.J. et al. (1999) Mol. Hum. Reprod. 5:1059.
  6. Vieira, P. et al. (1991) Proc. Natl. Acad. Sci. 88:1172.
  7. Hsu, D.-H. et al. (1990) Science 250:830.
  8. Goodman, R. et al. (1992) Biochem. Biophys. Res. Commun. 189:1.
  9. Windsor, W.T. et al. (1993) Biochemistry 32:8807.
  10. Syto, R. et al. (1998) Biochemistry 37:16943.
  11. Kotenko, S.V. et al. (1997) EMBO J. 16:5894.
  12. Kotenko, S.V. et al. (2000) J. Biol. Chem. 276:2725.
  13. Hor, S. et al. (2004) J. Biol. Chem. 279:33343.
  14. Sheppard, P. et al. (2003) Nat. Immunol. 4:63.
  15. Fitzgerald, D.C. et al. (2007) Nat. Immunol. 8:1372.
  16. Wu, K. et al. (2007) Cell. Mol. Immunol. 4:269.
  17. Blackburn, S.D. and E.J. Wherry (2007) Trends Microbiol. 15:143.

Long Name

Interleukin 10

Alternate Names

CSIF, GVHDS, IL10, IL10A, TGIF

Entrez Gene IDs

3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 102133450 (Cynomolgus Monkey); 100034187 (Equine); 493683 (Feline); 100715618 (Guinea Pig); 2949786 (Viral)

Gene Symbol

IL10

UniProt

Additional IL-10 Products

Product Documents for Mouse/Rat IL‑10 Antibody

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/Rat IL‑10 Antibody

For research use only

Citations for Mouse/Rat IL‑10 Antibody

Customer Reviews for Mouse/Rat IL‑10 Antibody (5)

4.6 out of 5
5 Customer Ratings
5 Stars
80%
4 Stars
0%
3 Stars
20%
2 Stars
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1 Stars
0%

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Customer Images


Showing  1 - 5 of 5 reviews Showing All
Filter By:
  • Mouse/Rat IL-10 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Myeloid cells
    Species: Mouse
    Verified Customer | Posted 08/28/2022
    Try this for flow, the signal is not too strong, may need optimize.
    Mouse/Rat IL‑10 Antibody AF519
  • Mouse/Rat IL-10 Antibody
    Name: Alex O'Neal
    Application: Western Blot
    Sample Tested: Bronchoalveolar lavage
    Species: Rat
    Verified Customer | Posted 11/30/2020
    We had an issue with shipping due to COVID, that caused the antibody to sit at room temperature for too long and become unusable. Fortunately we placed another order of the exact same product number, and it performed perfectly. I've included the photo of the gel after the shipping mishap
    Mouse/Rat IL‑10 Antibody AF519
  • Mouse/Rat IL-10 Antibody
    Name: Alex O'Neal
    Application: Western Blot
    Sample Tested: Control
    Species: Rat
    Verified Customer | Posted 11/30/2020
    This image was purely a test with an IL-10 control protein at different concentrations. Initially we were unable to detect the IL-10 protein in our samples, so we were testing to ensure that this antibody was functional. Turns out it was!
    Mouse/Rat IL‑10 Antibody AF519
  • Mouse/Rat IL-10 Antibody
    Name: Danlee Enzler
    Application: Immunohistochemistry
    Sample Tested: Spleen tissue
    Species: Mouse
    Verified Customer | Posted 04/24/2018
  • Mouse/Rat IL-10 Antibody
    Name: Anonymous
    Application: Block/Neutralize
    Sample Tested: Splenic B cells
    Species: Mouse
    Verified Customer | Posted 07/27/2016

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