Human IL‑10 Antibody

Name: Human IL‑10 Antibody
Brand: R&D Systems
SKU: AF-217-NA
Price: 189 USD
Availability: InStock
Rating: 4 (2 reviews)

R&D Systems | Catalog # AF-217-NA

(2)

Citations (39)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Primate - Macaca mulatta (Rhesus Macaque)

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Assay Development, Functional Assay, Mass Cytometry, Imaging Mass Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all IL-10 Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL-10
Ser19-Asn178
Accession # P22301

Specificity

Detects human IL-10 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant Epstein-Barr virus IL-10 is observed and less than 20% cross-reactivity with recombinant mouse IL‑10, recombinant feline IL-10, recombinant porcine IL‑10, recombinant equine IL-10, and recombinant rat 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 Human IL‑10 Antibody

Detection of Human IL‑10 by Western Blot.

Western blot shows conditioned media of HEK293 human embryonic kidney cell line either mock transfected or transfected with human IL-10. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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 1.

IL‑10 in Human Tonsil.

IL-10 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

IL‑10 in Human Tonsil.

IL-10 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human IL‑10 by Simple Western^TM.

Simple Western lane view shows conditioned media of HEK293 human embryonic kidney cell line either mock transfected or transfected with human IL-10, loaded at 0.2 mg/mL. A specific band was detected for IL-10 at approximately 23 kDa (as indicated) using 10 µg/mL of Goat Anti-Human IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

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

Recombinant Human IL-10 (Catalog # 217-IL) stimulates proliferation in the MC/9-2 mouse mast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-10 (5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA). The ND₅₀ is typically 0.2-1.0 µg/mL.

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Applications for Human IL‑10 Antibody

Application

Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Simple Western

10 µg/mL
Sample: HEK293 human embryonic kidney cell line transfected with human IL-10

Western Blot

1 µg/mL
Sample: HEK293 human embryonic kidney cell line transfected with human IL-10

Neutralization

Measured by its ability to neutralize IL‑10-induced proliferation in the MC/9‑2 mouse mast cell line. The Neutralization Dose (ND₅₀) is typically 0.2-1.0 µg/mL in the presence of 5 ng/mL Recombinant Human IL‑10.

Reviewed Applications

Read 2 reviews rated 4 using AF-217-NA in the following applications:

Immunohistochemistry (1 Review)
Immunohistochemistry-Frozen (1 Review)

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.

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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, IL-24, and IL-26/AK155 (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 human IL-10 shares 72%‑86% amino acid sequence identity with bovine, canine, equine, feline, mouse, ovine, porcine, and rat 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 (6, 8, 9). 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-10 R2 chains and activation of a signaling cascade involving JAK1, TYK2, and STAT3 (10). 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 (11‑13). IL-10 is a critical molecule in the control of viral infections and allergic and autoimmune inflammation (14‑16). It promotes phagocytic uptake and Th2 responses but suppresses antigen presentation and Th1 proinflammatory responses (2).

References

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

P22301

Additional IL-10 Products

Product Documents for Human IL‑10 Antibody

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Product Specific Notices for Human IL‑10 Antibody

For research use only

Related Research Areas

B Cell Development and Differentiation

Conventional/Classical Dendritic Cells

Cytokines and Receptors in Allergy and Asthma

Cytokines and Receptors in Angiogenesis

Cytokines and Receptors Research

gamma delta T Cells

Inflammatory Mediators

Isolation, Culture, and Identification of Natural Killer Cells

Macrophage Activation Markers

Microglia Activation Markers

Monocyte Markers

Myeloid-derived Suppressor Cells (MDSC)

Natural Killer T (NKT) Cells

Plasmacytoid Dendritic Cells

Regulatory T Cells (Tregs)

T Cell Cytokine Signaling

Citations for Human IL‑10 Antibody

Customer Reviews for Human IL‑10 Antibody (2)

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Human IL-10 Antibody

Name: Tyler Crowe

Application: Immunohistochemistry-Frozen

Sample Tested: Frozen human skin sections

Species: Human

Verified Customer | Posted 12/14/2018

Goat anti-IL10 (1:50, green) stained normal human skin, counterstained with DAPI (blue)

Block with 5% BSA, incubated with antibody 1:50 in blocking solution at 37C for 1 hour
Human IL-10 Antibody

Name: Stephanie Zandee

Application: Immunohistochemistry

Sample Tested: Adult brain

Species: Human

Verified Customer | Posted 04/13/2018

Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287 Used at 10ug/ml. Briefly, frozen brain sections were fixed in 4% PFA (Fisher Scientific), followed by antigen retrieval using heating in acid citric buffer (Vector, Burlingame, CA, USA). Endogenous avidin-biotin was blocked for 15 min (Vector). Sections were incubated with 10% horse serum in PBS (Biosera, Boussens, France) and Fc Receptor Blocking Solution was added (Human TruStain FcX Biolegend, London, UK). Primary antibody was added overnight at 4 °C and detected with donkey anti-goat-biotin (ab6578, Abcam), followed by streptavidin-alkaline phosphatase (SA-5100, Vector) and visualised with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector).