Human Total Serpin E1/PAI-1 Quantikine ELISA Kit

R&D Systems | Catalog # DTSE100

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL)

Sensitivity

0.046 ng/mL

Assay Range

0.313-20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
View all Serpin E1/PAI-1 ELISA Kits »

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

The Quantikine Human Total Serpin E1/PAI-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Serpin E1 in cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human Serpin E1 and has been shown to accurately quantitate the recombinant factor free and in complex with uPA, tPA, or vitronectin. Results obtained using natural human Serpin E1 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Serpin E1.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human Total Serpin E1 (active, latent, uPA, tPA, and Vitronectin complexed Serpin E1).

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.

Cell Culture Supernates, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 1.61 5.05 11.1 1.82 5.28 10.0
Standard Deviation 0.097 0.330 0.495 0.196 0.451 0.604
CV% 6.0 6.5 4.5 10.8 8.5 6.0

Recovery for Human Total Serpin E1/PAI-1 Quantikine ELISA Kit

The recovery of human Total Serpin E1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 105 90-119
Citrate Plasma (n=4) 104 87-118
EDTA Plasma (n=4) 99 85-113
Heparin Plasma (n=4) 103 81-116
Serum (n=4) 104 93-116

Linearity

To assess the linearity of the assay, samples containing high concentrations of human Serpin E1 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. All samples were diluted prior to assay.

Human Total Serpin E1/PAI-1 ELISA Linearity

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

Human Total Serpin E1/PAI-1 ELISA Standard Curve

Human Total Serpin E1/PAI-1 ELISA Standard Curve

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 Quantikine ELISA Kit

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.

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

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human Total Serpin E1/PAI-1 Quantikine ELISA Kit

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Protocols

View specific protocols for Human Total Serpin E1/PAI-1 Quantikine ELISA Kit (DTSE100):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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FAQs

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Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail