Human Total Serpin E1/PAI-1 Quantikine ELISA Kit

Catalog #: DTSE100
3 Citations Datasheet / COA / SDS

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DTSE100

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QC209: Quantikine Immunoassay Control Set 822 Human Total PAI-1

Human Total Serpin E1/PAI-1 ELISA Standard Curve

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All Serpin E1/PAI-1 ELISAs

Product Details

Procedure

Citations (3)

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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

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

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

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

Sensitivity

0.046 ng/mL

Assay Range

0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

	Intra-Assay Precision	Inter-Assay Precision
Sample	3	1	2	3
n	20	20	20	20
Mean (ng/mL)	11.1	1.82	5.28	10
Standard Deviation	0.495	0.196	0.451	0.604
CV%	4.5	10.8	8.5	6

Recovery

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.

Scientific Data

N/A

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Human Total Serpin E1/PAI-1 ELISA Standard Curve

Product Datasheets

Product Datasheet

COA

SDS

Preparation and Storage

Shipping

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

Storage

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

Background: Serpin E1/PAI-1

The human serpin superfamily consists of at least 35 members that target not only serine proteases, but also selected cysteine proteases and non-protease proteins. Serpins bind the protease active site resulting in a major conformational rearrangement that traps the enzyme in a covalent acyl-enzyme intermediate. As protease inhibitors, serpins have an array of functions including regulating blood clotting, the complement pathway, extracellular matrix remodeling, and cell motility. They are also involved in activities that extend beyond their ability to inhibit proteases. For instance, they may also regulate blood pressure, angiogenesis, or act as storage/transport proteins.

Long Name:

Plasminogen Activator Inhibitor

Entrez Gene IDs:

5054 (Human); 18787 (Mouse)

Alternate Names:

Endothelial plasminogen activator inhibitor; Nexin; PAI1; PAI-1; PAI1PAI-1; PAISerpin E1; PLANH1; PLANH1plasminogen activator inhibitor 1; serine (or cysteine) proteinase inhibitor, clade E (nexin, plasminogenactivator inhibitor type 1), member 1; Serpin E1; serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitortype 1), member 1

Related Research Areas

⚠ 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.

Assay Procedure

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.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

50 µL Assay Diluent

Add 50 µL of Assay Diluent to each well.

50 µL Standard, Control, or Sample

Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

200 µL Conjugate

Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 4 times.

200 µL Substrate Solution

Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

50 µL Stop Solution

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.

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

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Oxidative stress-induced endothelial dysfunction and decreased vascular nitric oxide in COVID-19 patients
Authors: V Montiel, I Lobysheva, L Gérard, M Vermeersch, D Perez-Morg, T Castelein, JB Mesland, P Hantson, C Collienne, D Gruson, MA van Dievoe, A Persu, C Beauloye, M Dechamps, L Belkhir, A Robert, M Derive, PF Laterre, AHJ Danser, X Wittebole, JL Balligand
EBioMedicine, 2022-02-23;77(0):103893.
Species: Human
Sample Types: Serum
Endurance Training Depletes Antioxidant System but Does Not Affect Endothelial Functions in Women with Abdominal Obesity: A Randomized Trial with a Comparison to Endurance-Strength Training
Authors: M Jamka, P Bogda?ski, P Krzy?anows, A Mi?kiewicz, J Karolkiewi, M Du?-?uchow, R M?dry, A Lisowska, A Gotz-Wi?ck, S Iskakova, J Walkowiak, E M?dry
Journal of Clinical Medicine, 2021-04-12;10(8):.
Species: Human
Sample Types: Whole Blood
Temporal trend of autonomic nerve function and HSP27, MIF and PAI-1 in type 1 diabetes
Authors: M Zimmerman, S Rolandsson, H Skärstrand, K Pourhamidi, A Gottsäter, P Wollmer, O Rolandsson, G Westergren, LB Dahlin
J Clin Transl Endocrinol, 2017-03-20;8(0):15-21.
Species: Human
Sample Types: Serum