Human Total Serpin E1/PAI-1 DuoSet ELISA

R&D Systems | Catalog # DY9387-05

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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

7.81-500 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human Total Serpin E1/PAI-1 DuoSet ELISA Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits
View all Serpin E1/PAI-1 ELISA Kits »

Product Summary for Human Total Serpin E1/PAI-1 DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human free and complexed Serpin E1 (Total Serpin E1). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, and plasma samples. Diluents for complex matrices, such as serum, and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human Total Serpin E1/PAI-1 DuoSet ELISA

Human Serpin E1 / PAI-1 ELISA Standard Curve

Human Serpin E1 / PAI-1 ELISA Standard Curve

Kit Contents for Human Total Serpin E1/PAI-1 DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Serpin E1/PAI-1

Serpin E1, also known as Plasminogen Activator Inhibitor-1 (PAI-1), is the primary inhibitor of urokinase-type and tissue-type plasminogen activators (uPA and tPA) which convert Plasminogen to Plasmin. The PA-Plasmin system is involved in fibrinolysis, tissue fibrosis, angiogenesis, wound healing, tumor cell invasion and metastasis, and obesity. Serpin E1 can promote clot formation, arterial re-endothelialization, and neointima formation while protecting against vascular intima thickening. It binds Vitronectin and inhibits vascular smooth muscle cell and endothelial cell adhesion, proliferation, and motility. Serpin E1 binds and induces the LRP-dependent internalization of cell surface complexes containing uPA, uPAR, and Integrin alpha V beta 3. Serpin E1 secretion is upregulated during inflammation, physical injury, and exposure to Angiotensin II.

Long Name

Plasminogen Activator Inhibitor

Alternate Names

Nexin, PAI-1, PAI1, PLANH1

Entrez Gene IDs

5054 (Human); 18787 (Mouse)

Gene Symbol

SERPINE1

Additional Serpin E1/PAI-1 Products

Product Documents for Human Total Serpin E1/PAI-1 DuoSet ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human Total Serpin E1/PAI-1 DuoSet ELISA

For research use only

Citations for Human Total Serpin E1/PAI-1 DuoSet ELISA

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Protocols

View specific protocols for Human Total Serpin E1/PAI-1 DuoSet ELISA (DY9387-05):

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL of Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
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