Human IL-6R alpha Antibody

Catalog #: MAB227R Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB227R-100
MAB227R-SP

Detection of IL‑6 R alpha in U937 Human Cell Line by Flow Cytometry.

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Human IL-6R alpha Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL-6 R alpha in ELISAs and Western blots. In ELISAs, no cross-reactivity or interference was observed with recombinant human(rh) IL-1 alpha, recombinant mouse (rm) IL-1 alpha, rhIL-1 beta, rmIL-1 beta, rhIL-1ra, rhIL-2, rhIL-3, rmIL-3, rhIL-4, rmIL-4, rhIL-5, rmIL-5, rhIL-6, rmIL-6, rhIL-7, rmIL-7, rhIL-8, rhIL-9, rmIL-9, rhIL-10, or rhIL-11.

Source

Recombinant Monoclonal Mouse IgG₁ Clone # 17506R

Purification

Protein A or G purified from cell culture supernatant

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑6 R alpha
Leu20-Asp339
Accession # P08887

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

1 µg/mL

Recombinant Human IL‑6 R alpha (Catalog # 227-SR)

Flow Cytometry

0.25 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Human IL-6R alpha Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Human IL-6R alpha Biotinylated Antibody (Catalog # BAF227)

Standard: Recombinant Human IL-6R alpha Protein (Catalog # 227-SR)

Neutralization

Measured by its ability to neutralize IL‑6 R alpha -mediated enhancement of IL‑6-dependent Inhibition of cell proliferation in the M1 mouse myeloid leukemia cell line. The Neutralization Dose (ND₅₀) is typically 0.04-0.2 µg/mL in the presence of 30 ng/mL Recombinant Human IL‑6 R alpha and 30 ng/mL Recombinant Human IL‑6.

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

Flow Cytometry

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Detection of IL‑6 R alpha in U937 Human Cell Line by Flow Cytometry. U937 human histiocytic lymphoma cell line was stained with Human IL-6 R alpha Monoclonal Antibody (Catalog # MAB227R, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgGF(ab')2 Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Neutralization

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IL-6 R alpha Enhancement of IL-6-dependent Inhibition of Cell Proliferation and Neutralization by Human IL-6 R alpha Antibody. Recombinant Human IL-6 Ra (Catalog # 227-SR) enhances Recombinant Human IL-6 (Catalog # 206-IL) inhibition of proliferation in the M1 mouse myeloid leukemia cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Enhancement of Recombinant Human IL-6 (30 ng/mL) activity elicited by Recombinant Human IL-6 Ra (30 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-6 Ra Monoclonal Antibody (Catalog # MAB227R). The ND₅₀ is typically 0.04-0.2 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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.

Background: IL-6R alpha

The multi-functional factor interleukin 6 (IL-6) exerts its activities through binding to a high-affinity receptor complex consisting of two membrane glycoproteins: an 80 kDa component receptor that binds IL-6 with low affinity (IL-6 R alpha ) and a signal-transducing component of 130 kDa (gp130) that does not bind IL-6 by itself, but is required for high-affinity binding of IL-6 by the complex. Both components of the receptor complex, IL-6 R alpha and gp130 have been cloned, sequenced, and expressed (1-4).

A soluble form of the IL-6 R alpha has been found in the urine of healthy adult humans (5). This soluble receptor apparently arises from proteolytic cleavage of membrane-bound IL-6 R alpha and is about 50kDa in size. No naturally-occurring mRNA encoding a truncated form of the IL-6 R alpha has been reported. Soluble forms of human and murine IL-6 R alpha s have been constructed, however, by insertion of termination codons into the regions of the IL-6 R alpha cDNAs encoding the external portions of the receptors and prior to the transmembrane domains. These soluble receptors have been expressed in COS-7 and CHO cells and have been shown to bind to IL-6 in solution and to augment the activity of IL-6 as a result of the binding of the IL-6/IL-6 R alpha complex to membrane-bound gp130 (6, 7).

References