Immunoprecipitaiton with the Human IL-10 Rα antibody (Catalog # AF-274-NA) followed by western blotting for Human IL-10 Rα produces a 150 kDa band. What is the identity of this band?

Name: Human IL-10 R alpha Antibody
Brand: R&D Systems
SKU: AF-274-NA
Price: 459 USD

Human IL-10 Rα antibody has not been validated for immunoprecipitaiton application. The MW of Human IL-10 Rα is about 90-110 kDa and the receptor binds with high affinity to the IL-10 homodimer, which has a MW of 40 kDa. Under non-denaturing conditions, the IL-10Rα may be complexed with IL-10 homodimer to give a 150 kDa band.

Human IL-10 R alpha Antibody

Catalog #: AF-274-NA
2 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF-274-NA
AF-274-SP

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Citations (2)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human IL-10 R alpha Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL‑10 R alpha in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse
IL-10 R alpha is observed and less than 5% cross-reactivity with recombinant human (rh) CNTF sR alpha and rhIL-2 sR gamma is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑10 R alpha
His22-Asn235
Accession # Q13651

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.

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human IL‑10 R alpha (Catalog # 274-R1)

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize IL‑10 R alpha -mediated IL‑10 response in LPS-activated human peripheral blood mononuclear cells (PBMC) [Ralph, P. et al. (1991) J. Immunology 148:808]. The Neutralization Dose (ND₅₀) is typically 20-40 µg/mL in the presence of 0.25 ng/mL Recombinant Human IL‑10 and 0.25-3 ng/mL LPS.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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IL‑10 R alpha in Human PBMCs. IL‑10 R alpha was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using 15 µg/mL Goat Anti-Human IL‑10 R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑274‑NA) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization

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IL‑10 Inhibition of IL‑1 beta secretion and Neutralization by Human IL‑10 R alpha Antibody. Recombinant Human IL-10 (Catalog # 217-IL) inhibits IL-1 beta secretion in LPS-activated human peripheral blood mononuclear cells (PBMC) in a dose-dependent manner (orange line), as measured by the Human IL-1 beta /IL-1F2 Quantikine ELISA Kit (Catalog # DLB50). IL-1 beta secretion inhibited by Recombinant Human IL-10 (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-10 Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-274-NA). The ND₅₀ is typically 20-40 µg/mL in the presence of LPS (0.25-3 ng/mL).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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.

Background: IL-10 R alpha

IL-10, initially designated cytokine synthesis inhibitory factor (CSIF), is a potent immunosuppressant of macrophage functions. IL-10 is also a pleiotropic cytokine with multiple immunostimulatory as well as immunosuppressive effects on a variety of other cell types. IL-10 binds specifically and with high affinity to cell-surface receptors. Mouse and human cDNA clones encoding the ligand-binding IL-10 receptor (IL-10 R) have been isolated. The IL-10 R mRNA has been detected in all cell types that are known to respond to IL-10.

Human and mouse IL-10 receptors are structurally related to the IFN-gamma receptor. These receptors are members of the class II subgroup of the cytokine receptor superfamily. The deduced amino acid sequence of human IL-10 R is approximately 60% identical to mouse IL-10 R. Although human IL-10 has cross-species activities and is active on mouse cells, mouse IL-10 is species-specific in its actions and does not bind to the human IL-10 receptor. The human IL-10 R gene has been mapped to chromosome 11q23.3. Recombinant IL-10 soluble receptor, consisting of the extracellular domain of IL-10 R, binds IL-10 with high affinity in solution and is a potent IL-10 antagonist.

Long Name

Interleukin 10 Receptor alpha

Entrez Gene IDs

3587 (Human); 16154 (Mouse); 102127536 (Cynomolgus Monkey)

Alternate Names

CD210 antigen; CDw210a; HIL-10R; IBD28; IL-10 R alpha; IL-10 receptor subunit alpha; IL10R alpha; IL-10R subunit 1; IL-10R1; IL10RA; IL-10Ra; IL10RIL-10R subunit alpha; interleukin 10 receptor, alpha; interleukin-10 receptor alpha chain; Interleukin-10 receptor subunit 1; interleukin-10 receptor subunit alpha

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Citations for Human IL-10 R alpha Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Multi-Step Regulation of the TLR4 Pathway by the miR-125a~99b~let-7e Cluster
Authors: Graziella Curtale, Tiziana A. Renzi, Massimiliano Mirolo, Lorenzo Drufuca, Manuel Albanese, Mariacristina De Luca et al.
Frontiers in Immunology
Differential regulation of CD44 expression by lipopolysaccharide (LPS) and TNF-alpha in human monocytic cells: distinct involvement of c-Jun N-terminal kinase in LPS-induced CD44 expression.
Authors: Gee K, Lim W, Ma W, Nandan D, Diaz-Mitoma F, Kozlowski M, Kumar A
J. Immunol., 2002-11-15;169(10):5660-72.
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

FAQs

Immunoprecipitaiton with the Human IL-10 Rα antibody (Catalog # AF-274-NA) followed by western blotting for Human IL-10 Rα produces a 150 kDa band. What is the identity of this band?
- Human IL-10 Rα antibody has not been validated for immunoprecipitaiton application. The MW of Human IL-10 Rα is about 90-110 kDa and the receptor binds with high affinity to the IL-10 homodimer, which has a MW of 40 kDa. Under non-denaturing conditions, the IL-10Rα may be complexed with IL-10 homodimer to give a 150 kDa band.

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