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.

Human IL-10 R alpha Antibody

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF-274-NA

Citations (1)

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Key Product Details

Species Reactivity:	Human
Immunogen:	S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑10 R alpha His22-Asn235 Accession # Q13651
Applications:	Block/Neutralize, Immunocytochemistry/Immunofluorescence, Western Blot
Label:	Unconjugated
Source:	Polyclonal Goat IgG
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.

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Applications for Human IL-10 R alpha Antibody

	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

Item	Value
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.

Scientific Data Examples for Human IL-10 R alpha Antibody

	Immunocytochemistry 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 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).

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Endotoxin level	<0.10 EU per 1 μg of the antibody by the LAL method.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.

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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.

Item	Value
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, IL-10R subunit 1, IL-10R1, IL-10Ra, IL10R alpha, IL10RA, IL10RIL-10R subunit alpha, Interleukin-10 receptor subunit 1, interleukin 10 receptor, alpha, interleukin-10 receptor alpha chain, interleukin-10 receptor subunit alpha

Citations for Human IL-10 R alpha Antibody (1)

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Citations are publications that use Bio-Techne products. Selected citations for Human IL-10 R alpha Antibody include:

Species: Human
Sample Types: Whole Cells
Applications: Neutralization

Gee K et al. (2002), 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. J. Immunol., 169(10):5660-72.
PMID: 12421945

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FAQs for Human IL-10 R alpha Antibody

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.

Human IL-10 R alpha Antibody

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Applications for Human IL-10 R alpha Antibody

Scientific Data Examples for Human IL-10 R alpha Antibody

Immunocytochemistry

Neutralization

Formulation, Preparation and Storage

Reconstitution Calculator

Background: IL-10 R alpha

Citations for Human IL-10 R alpha Antibody (1)

FAQs for Human IL-10 R alpha Antibody

Product Documents for Human IL-10 R alpha Antibody

Product Datasheets

COA

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