Human IL-6R alpha (Research Grade Tocilizumab Biosimilar) Antibody

Catalog #: MAB11482 Datasheet / COA / SDS

Recombinant Monoclonal Antibody For Research Use Only

Catalog #	Availability	Size / Price	Qty
MAB11482-100
MAB11482-SP

Detection of IL-6R alpha in HEK293 cells transfected with hIL-6Ra and eGFP by Flow Cytometry

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Human IL-6R alpha (Research Grade Tocilizumab Biosimilar) Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL-6R alpha in direct ELISAs.

Source

Recombinant Monoclonal Human IgG₁ Clone # Hu203

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human IL-6R alpha
Arg387-Arg468
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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

ELISA

This antibody functions as an ELISA detection antibody for the specific antigen in direct ELISA. Colorimetric detection is performed after addition of a suitable substrate.

Flow Cytometry

0.25 µg/10⁶ cells

hIL-6Ra HEK/eGFP transfectants

Blockade of Receptor-ligand Interaction

0.25 µg/10⁶ cells

Blocking assay in hIL-6Ra HEK/eGFP transfecants (see details below)

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-6R alpha in HEK293 cells transfected with hIL-6Ra and eGFP by Flow Cytometry HEK293 cells transfected with hIL-6Ra and eGFP were stained with Human Anti-Human IL-6R alpha (Research Grade Tocilizumab Biosimilar) Monoclonal Antibody (Catalog # MAB11482, filled histogram) followed by Allophycocyanin-conjugated Anti-Human IgG Secondary Antibody (Catalog # F0135) or unstained cells (open blue histogram). View our protocol for Staining Membrane-associated Proteins.

Blockade of Receptor-ligand Interaction

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IL-6 binding to IL-6Ra transfected HEK293 Human Cell line is Blocked by Human IL-6Ra (Tocilizumab) Antibody. In a functional flow cytometry test, biotinylated recombinant IL-6 (50ng/mL, BT7270B) binds to HEK293 human embyonic kidney cell line transfected with human IL-6Ra and eGFP. (A) Binding is blocked by 25 ug/mL of Human anti-Human IL-6Ra Monoclonal Antibody (Catalog# MAB11482) or (B) Protein binding only. Protein binding was detected by staining cells with Streptavidin-APC (F0050)

ELISA

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Human IL-6R alpha ELISA Standard Curve Direct ELISA binding curve demonstrating the recognition of Human Anti-Human IL-6R alpha (Research Grade Tocilizumab Biosimilar) Monoclonal Antibody (Catalog # MAB11482) to IL-6R alpha. The target protein was coated onto the microplate well surface, followed by binding of the antibody. A goat anti-human HRP conjugate was used for detection.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 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-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 (1‑4). Within the extracellular domain, human IL‑6 R alpha shares 52% aa sequence identity with
mouse and rat IL‑6 R alpha.

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. 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