Human IL‑1 beta /IL‑1F2 Antibody

Name: Human IL‑1 beta /IL‑1F2 Antibody
Brand: R&D Systems
SKU: MAB601R
Availability: InStock

R&D Systems | Catalog # MAB601R

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Product Documents

Recombinant Monoclonal Antibody.

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Human

Applications

Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization, Immunocytochemistry, Simple Western

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG₁ Clone # 2805R

View all IL-1 beta/IL-1F2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human IL-1 beta /IL-1F2

Specificity

Detects human IL-1 beta /IL-1F2 in sandwich ELISAs and Western blots. In sandwich ELISAs, less than 4% cross-reactivity with recombinant rat (rr) IL‑1 beta and less than 0.1% with recombinant porcine (rp) IL-1 beta, recombinant human IL-1 alpha, rpIL-1 alpha, rrIL-1 alpha, recombinant mouse (rm) IL-1 alpha, and rmIL-1 beta is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Endotoxin Level

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

Scientific Data Images for Human IL‑1 beta /IL‑1F2 Antibody

Detection of Human IL‑1 beta /IL‑1F2 by Western Blot.

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS for 3 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB601R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL-1 beta /IL-1F2 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of IL‑1 beta /IL‑1F2 in THP‑1 Human Cell Line.

IL‑1 beta /IL‑1F2 was detected in immersion fixed THP‑1 human acute monocytic leukemia cell line using Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB601R) at 8 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the cytoplasm of THP-1 cells treated with 200nM PMA for 24 hours then 10ug/mL LPS for 24 hours. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

IL‑1 beta /IL‑1F2 in Human PBMCs.

IL-1 beta /IL-1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with 1 µg/mL LPS and 3 µM monensin for 24 hours using Mouse Anti-Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB601R) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human IL‑1 beta /IL‑1F2 by Simple Western^TM.

Simple Western lane view shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 200 nM and 10 μg/mL LPS for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 37 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB601R). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Cell Proliferation Induced by IL-1 beta /IL-1F2 and Neutralization by Human IL-1 beta /IL-1F2 Antibody.

Recombinant HumanIL-1 beta /IL-1F2 (Catalog # 201-LB) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL-1 beta /IL-1F2 (50 pg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-1 beta /IL-1F2 Monoclonal Antibody (Catalog # MAB601R). The ND₅₀ is typically 50-200 ng/mL.

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Applications for Human IL‑1 beta /IL‑1F2 Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with 1 μg/mL LPS and 3 μM monensin for 24 hours and THP-1 human acute monocytic leukemia cell line untreated and treated with PMA and LPS

Simple Western

10 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line treated with PMA and LPS

Western Blot

1 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line treated with PMA and LPS

Neutralization

Measured by its ability to neutralize IL‑1 beta /IL‑1F2-induced proliferation in the D10.G4.1 mouse helper T cell line. The Neutralization Dose (ND50) is typically 50-200 ng/mL in the presence of 50 pg/mL Recombinant Human IL‑1 beta /IL‑1F2.

Human IL-1 beta /IL-1F2 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)

Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:

Detection Reagent: Human IL‑1 beta /IL‑1F2 Biotinylated Antibody (Catalog # BAF201)
Standard: Recombinant Human IL-1 beta/IL-1F2 Protein (Catalog # 201-LB)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 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. See Certificate of Analysis for details.
*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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: IL-1 beta/IL-1F2

IL-1 is a name that designates two pleiotropic cytokines, IL-1 alpha (IL-1F1) and IL-1 beta (IL-1F2, IL1B), which are the products of distinct genes. IL-1 alpha and IL-1 beta are structurally related polypeptides that share approximately 21% amino acid (aa) identity in human. Both proteins are produced by a wide variety of cells in response to inflammatory agents, infections, or microbial endotoxins. While IL-1 alpha and IL-1 beta are regulated independently, they bind to the same receptor and exert identical biological effects. IL-1 RI binds directly to IL-1 alpha or IL-1 beta and then associates with IL-1 R accessory protein (IL-1 R3/IL-1 R AcP) to form a high-affinity receptor complex that is competent for signal transduction. IL-1 RII has high affinity for IL-1 beta but functions as a decoy receptor and negative regulator of IL-1 beta activity. IL-1ra functions as a competitive antagonist by preventing IL-1 alpha and IL-1 beta from interacting with IL-1 RI. Intracellular cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. The 17 kDa molecular weight mature human IL-1 beta shares 96% aa sequence identity with rhesus and 67%-78% with canine, cotton rat, equine, feline, mouse, porcine, and rat IL-1 beta. IL-1 beta functions in a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. IL-1 beta dysregulation is implicated in many pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases.