PAI-1 ELISA Kit, Human

Catalog number: KHC3071

Novex™  Related applications: Protein Assays and Analysis

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Description

The Human Plasminogen Activator Inhibitor-1 (PAI-1) ELISA research-use-only kit is to be used for the quantitative determination of human PAI-1 in samples (e.g., serum, EDTA and heparin plasma, cell culture supernatants, etc.) using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant human PAI-1.

Performance characteristics
• Sensitivity: <30 pg/mL
• Standard curve range: 62.5–4,000 pg/mL
• Sample types: serum, plasma (EDTA, heparin), cell culture supernatants
• Species cross-reactivity: human
• Sample volume: 100 μL (1:2)
• Total assay incubation time: 5 hrs

Principle of the method
The Human PAI-1 kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal antibody specific for Human PAI-1 has been coated onto the wells of the microtiter strips provided. During the first incubation, standards of known human PAI-1 content, controls, and unknown samples are pipetted into the coated wells. After washing, polyclonal biotinylated second antibody, is added. After washing, streptavidin-peroxidase (enzyme) is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of Human PAI-1 present in the original specimen.

Background information
Plasminogen activator inhibitor-1 (PAI-1) is the principal inhibitor of tissue-type and urokinase-type plasminogen activators (tPA and uPA), which convert plasminogen to plasmin. PAI-1 is a member of serine protease inhibitor superfamily, also known as Serpin E1. PAI-1 inhibits the serine proteases tPA and uPA/urokinase. It has been also demonstrated that PAI-1 activates the JAK/Stat signaling pathway and stimulates cell migration by binding to the low-density lipoprotein receptor-related protein.

PAI-1 is synthesized and secreted by many tissue and cell types. Free PAI-1 is relatively unstable in its active form and readily converts into a latent, inactive form by spontaneous insertion of its reactive center loop into the β-sheet core of the protein. It was demonstrated that binding of vitronectin to PAI-1 could stabilize the active form by preventing the conformation change. The active form of PAI-1 binds tightly with uPA and tPA in a 1:1 ratio. After the formation of an initial docking complex, the proteases cleave the reactive center loop of PAI-1 to form a stable, covalent complex, resulting in the inactivation of the targeted serine protease.

Each ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See product insert for more information on validation.

Related links
Learn more about ELISA kits
Learn more about other immunoassays
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Gene Aliases: clade E nexin, plasminogen activator inhibitor type 1, member 1, PAI, PAI-1, PAI1, PLANH1
Protein Subtype: Protease Inhibitors
Accession Number: NM_000602.4, NP_000593.1, NP_001158885.1, NM_001165413.2
Shipping Condition: Wet Ice
Marker: PAI-1
Gene ID: 5054
Species: Human, Rat
Gene Symbol: SERPINE1
Sensitivity: <30 pg/mL
Target Gene: serpin peptidase inhibitor
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 assays
Concentration: 62.5-4000 pg⁄ml
Sample Volume: 100 µl
Incubation Time: 5 hrs
Detection Method: Colorimetric
Research Category: Immunology
Sample Type (Specific): Plasma, Serum, Supernatants
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