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IL-1 alpha ELISA Kit, Human

Catalog number: KAC1191

Novex™  Related applications: Protein Assays and Analysis

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Description

The Human Interleukin-1 alpha (IL-1 alpha, IL1A) ELISA research-use-only kit is to be used for the quantitative determination of IL-1 alpha in samples (see sample types indicated) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant forms of this target.

Performance characteristics

• Sensitivity: <1 pg/mL
• Standard curve range: 3.9‒250 pg/mL
• Sample type(s): serum, plasma, cell culture supernatant
• Specificity: natural and recombinant human IL-1 alpha
• Cross-reactivity: see kit manual for cross-species and/or cross-target reactivity
• Sample volume: 50 μL
• Total assay time: 3 hours

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Principle of the method

The human IL-1 alpha solid-phase sandwich ELISA (enzyme-linked immunosorbent assay) is designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody has been pre-coated in the wells of the supplied microplate. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. The sandwich is formed by the addition of the second (detector) antibody, binding to the target on a different epitope from the capture antibody. A conjugated enzyme has been incorporated into the assay. After incubation and washing steps to rid the microplate of unbound substances, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Target information

Human interleukin-1 (IL-1) is a key mediator of the host response to various infectious, inflammatory and immunologic challenges. Two distinct polypeptides, IL-1α and IL-1β, mediate the biological activities and bind to the same cell surface receptors. Both are initially synthesized as 31 kDa precursors that are subsequently found as 17 kDa mature proteins. A large proportion of IL-1α precursor is retained intracellularly. The 31 kDa membrane-bound form of IL-1α has also been reported to be present on the cell surfaces. This membrane-bound IL-1α acts biologically in a paracrine fashion on those adjacent cells having IL-1 receptors. Intracellular IL-1β consists exclusively of the 31 kDa precursor form that shows little or no biological activity in comparison to the 17 kDa processed form.

Two distinct IL-1 receptors, IL-1 receptors type I and II (IL-1 RI and IL-1 RII) have been cloned and characterized. Both type I and type II receptors are members of the immunoglobulin superfamily. Human IL-1 RI is an 80 kDa transmembrane protein with 552 amino acids. It has a single 22 amino acid transmembrane region and a long cytoplasmic tail of 212 amino acids, which participates in signal transduction. Type 1 receptors are found on most cells, including T cells, endothelial cells, hepatocytes, fibroblasts and keratinocytes. The type II 60 kDa receptor is similar to the type 1 receptor in its extracellular and transmembrane regions. IL-1 RII has a cytoplasmic domain of only 29 amino acids and is incapable of signaling. IL-1 RII is the predominant receptor on B cells, monocytes and neutrophils. The two types of receptors show differential binding characteristics for IL-1α and IL-1β. In general, IL-1α binds better to the type I receptor and IL-1β binds better to the type II receptor. Soluble forms of IL-1 RI and IL-1 RII have been reported in sera. In addition, a receptor accessory protein (IL-1R-AcP) has been identified. This accessory protein forms a complex with IL-1 RI after it is bound by either IL-1α or IL-1β and increases the binding affinity of this receptor for IL-1.

The primary sources of IL-1 are blood monocytes and tissue macrophages. Other specialized cells such as T and B cells, various epithelial, endothelial and some mesenchymal cells can also produce IL-1. IL-1β is the major form secreted by monocytes and macrophages which are believed to be the main source of circulating (plasma) IL-1. Inhibition of IL-1 activity has been described in plasma and other biological fluids. IL-1 affects several unrelated tissues and is a main mediator of the "acute phase" inflammatory responses characterized by alterations in metabolic, endocrine and immunologic functions. This cytokine has an essential role in T cell activation, providing one of the necessary signals for IL-2 (T cell growth factor) production. It is the main mediator of inflammatory processes by acting on the nervous system (fever, sleep, anorexia), on bone marrow-derived cells (chemotaxis and/or activation of neutrophils, monocytes and lymphocytes) and on various tissues (fibroblast proliferation, resorption of cartilage and bone matrices, glial cell proliferation, stimulation of endothelial cell procoagulant activity, etc.). Most of these activities are directly attributable to IL-1β, but others are mediated in collaboration with other cytokines such as IL-6, interferons, and tumor necrosis factor. IL-1 stimulates the production or acts synergistically with these cytokines and the final biological activity is thus the result of a network of interactions between these various mediators.

Related links

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For Research Use Only. Not for use in diagnostic procedures.

Specifications

Gene Aliases: IL-1A, IL1, IL1-ALPHA, IL1F1
Accession Number: NP_000566.3, NM_000575.3
Shipping Condition: Wet Ice
Marker: IL-1α
Gene ID: 3552
Species: Human
Gene Symbol: IL1A
Sensitivity: 1 pg/ml
Target Gene: interleukin 1, alpha
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 assays
Concentration: 3.9-250 pg⁄ml
Sample Volume: 50 µl
Protein Family: Cytokines & Receptors
Incubation Time: 3 hrs
Detection Method: Colorimetric
Research Category: Immunology
Sample Type (Specific): Plasma, Supernatants, Serum
For Use With (Equipment): Microplate Reader

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.

Documents

Manuals & protocols

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