Human IL-1 beta /IL-1F2 Antibody

Catalog #: MAB601R Datasheet / COA / SDS

Recombinant Monoclonal Antibody

Catalog #	Availability	Size / Price	Qty
MAB601R-100
MAB601R-SP
MAB601R-500
MAB601R-01M

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

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Human IL-1 beta /IL-1F2 Antibody Summary

Species Reactivity

Human

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.

Source

Recombinant Monoclonal Mouse IgG₁ Clone # 2805R

Purification

Protein A or G purified from cell culture supernatant

Immunogen

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

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

See below

Simple Western

10 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

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

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Human IL‑1 beta /IL‑1F2 Biotinylated Antibody (Catalog # BAF201)

Standard: Recombinant Human IL-1 beta/IL-1F2 Protein (Catalog # 201-LB)

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.

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

Western Blot

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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.

Immunocytochemistry

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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.

Simple Western

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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.

Neutralization

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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.

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-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.

Long Name

Interleukin 1 beta

Entrez Gene IDs

3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)

Alternate Names

catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta