Detects human IL‑1ra/IL-1F3 in direct ELISAs and Western blots. In direct ELISAs and Western blots (non-reducing conditions), less than 15% cross-reactivity with recombinant mouse IL-1ra 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-1ra/IL-1F3 Antibody

Detection of Human IL‑1ra/IL‑1F3 by Western Blot.

Western blot shows lysates of RT-4 human bladder carcinoma cell line and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human IL-1ra/IL-1F3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-280-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-1ra/IL-1F3 at approximately 18-20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

IL‑1ra/IL‑1F3 in Human Skin.

IL-1ra/IL-1F3 was detected in immersion fixed paraffin-embedded sections of human skin using Goat Anti-Human IL-1ra/IL-1F3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-280-NA) at 5 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). Specific staining was localized to keratinocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human and Mouse IL‑1ra/IL‑1F3 by Simple Western^TM.

Simple Western lane view shows lysates of RT-4 human bladder carcinoma cell line, A431 human epithelial carcinoma cell line, human skin cancer tissue, and mouse skin cancer tissue, loaded at 0.2 mg/mL. A specific band was detected for IL-1ra/IL-1F3 at approximately 24-26 kDa (as indicated) using 20 µg/mL of Goat Anti-Human IL-1ra/IL-1F3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-280-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.

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Applications for Human IL-1ra/IL-1F3 Antibody

Application

Recommended Usage

Immunohistochemistry

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

Simple Western

20 µg/mL
Sample: RT-4 human bladder carcinoma cell line, A431 human epithelial carcinoma cell line, human skin cancer tissue, and mouse skin cancer tissue

Western Blot

0.5 µg/mL
Sample:

RT-4 human bladder carcinoma cell line and A431 human epithelial carcinoma cell line

Neutralization

Measured by its ability to neutralize IL‑1ra/IL‑1F3 inhibition of IL‑1 alpha/IL‑1F1-dependent proliferation in the D10.G4.1 mouse helper T cell line. Symons, J.A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 272. The Neutralization Dose (ND₅₀) is typically ≤4 µg/mL in the presence of 50 ng/mL Recombinant Human IL‑1ra/IL‑1F3 and 50 pg/mL Recombinant Human IL‑1 alpha/IL‑1F1.

Reviewed Applications

Read 2 reviews rated 5 using AF-280-NA in the following applications:

ELISA (2 Reviews)

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.

Background: IL-1ra/IL-1F3

IL-1ra was originally isolated from the urine of patients with monocytic leukemia and has also been purified from adherent monocytes. The naturally occurring, fully glycosylated form has an apparent molecular weight of about 25,000 Daltons. The protein shows 26% amino acid homology to IL-1 beta and 19% homology to IL-1 alpha. It will compete with either factor for receptor binding, but does not interact with either one. Human IL-1ra will bind to both types of IL-1 receptor (I and II) on human cells, but reportedly will not block binding to the type II receptor on murine pre-B cell lines. The recombinant, non-glycosylated form of IL-1ra blocks binding of IL-1 to its receptor equally as well as the naturally-occurring, glycosylated form. The IL-1ra has been shown to block the inflammatory responses induced by IL-1 both in vitro and in vivo. Currently, pre-clinical and clinical studies are underway to test possible therapeutic applications for IL-1ra in the treatment of sepsis, rheumatoid arthritis and chronic myelogenous leukemia.

Long Name

Interleukin 1 Receptor Antagonist

Alternate Names

DIRA, ICIL-1ra, IL-1F3, IL-1ra3, IL-1RN, IL1ra, IL1RN, MVCD4

Entrez Gene IDs

3557 (Human); 16181 (Mouse); 60582 (Rat); 397499 (Porcine); 100034236 (Equine)

Gene Symbol

IL1RN

UniProt

P18510

Additional IL-1ra/IL-1F3 Products

Product Documents for Human IL-1ra/IL-1F3 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human IL-1ra/IL-1F3 Antibody

For research use only

Citations
Reviews

Human IL-1ra/IL-1F3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF-280-NA

Key Product Details

Species Reactivity

Validated:

Cited:

Applications