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Human eNOS Quantikine ELISA Kit

Catalog #: DEN00
21 Citations Datasheet / COA / SDS

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Citations (21)
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Summary Precision Linearity Product Datasheets Preparation and Storage Background Related Research Areas

Human eNOS Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Lysates (100 uL)
Sensitivity
25 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Lysates)
Specificity
Natural and recombinant human eNOS
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity not tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human eNOS Immunoassay kit is a 4.5 hour solid phase ELISA designed to measure eNOS from endothelial cells. It contains E. coli-expressed recombinant human eNOS and antibodies raised against the recombinant enzyme. It has been shown to accurately quantitate recombinant human eNOS. Results obtained using natural human eNOS showed linear curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human eNOS.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Lysates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 127 619 1280 129 621 1275
Standard Deviation 6.2 22.9 51.8 9.6 27.5 45.7
CV% 4.9 3.7 4 7.4 4.4 3.6

Linearity

To assess the linearity of the assay, five cell lysate samples containing high concentrations of eNOS were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

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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 -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: eNOS

Nitric oxide (NO) is a pleiotropic signaling molecule implicated in diverse biological processes including inhibition of platelet aggregation, regulation of neurotransmission, vasodilation, immune responses, and inflammation. NO is synthesized from L-arginine and O2 by three nitric oxide synthase (NOS) enzymes termed endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS (iNOS). Each enzyme isoform is expressed in a variety of tissues and cell types. While eNOS and nNOS generally exhibit constitutive expression and are involved in physiological signaling and maintenance functions, iNOS expression is induced by inflammatory stimuli and is associated with both normal and pathological immune responses.

Long Name:
Endothelial Nitric Oxide Synthase
Entrez Gene IDs:
4846 (Human); 18127 (Mouse)
Alternate Names:
Constitutive NOS; EC 1.14.13.39; ECNOS; Endothelial NOS; endothelial; eNOS; eNOScNOS; nitric oxide synthase 3 (endothelial cell); NOS type III; NOS3; NOSIII

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.
  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. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 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 on the shaker.
  7.   Aspirate and wash 3 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. 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.

Citations for Human eNOS 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.

21 Citations: Showing 1 - 10
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  1. Toward engineering a human neoendothelium with circulating progenitor cells.
    Authors: Allen JB, Khan S, Lapidos KA, Ameer GA
    Stem Cells, 2010-02-01;28(2):318-28.
    Species: Human
    Sample Types: Cell Lysates
  2. Intradermal delivery of plasmid VEGF(165) by electroporation promotes wound healing.
    Authors: Ferraro B, Cruz YL, Coppola D, Heller R
    Mol. Ther., 2009-02-24;17(4):651-7.
    Species: Rat
    Sample Types: Tissue Homogenates
  3. Smoking cessation early in pregnancy and birth weight, length, head circumference, and endothelial nitric oxide synthase activity in umbilical and chorionic vessels: an observational study of healthy singleton pregnancies.
    Authors: Andersen MR, Simonsen U, Uldbjerg N, Aalkjaer C, Stender S
    Circulation, 2009-02-02;119(6):857-64.
    Species: Human
    Sample Types: Cell Lysates
  4. Statins restore ischemic limb blood flow in diabetic microangiopathy via eNOS/NO upregulation but not via PDGF-BB expression.
    Authors: Fujii T, Onimaru M, Yonemitsu Y, Kuwano H, Sueishi K
    Am. J. Physiol. Heart Circ. Physiol., 2008-04-25;294(6):H2785-91.
    Species: Human, Mouse
    Sample Types: Cell Lysates, Tissue Homogenates
  5. Hydroxyurea nitrosylates and activates soluble guanylyl cyclase in human erythroid cells.
    Authors: Cokic VP, Andric SA, Stojilkovic SS, Noguchi CT, Schechter AN
    Blood, 2007-11-09;111(3):1117-23.
    Species: Human
    Sample Types: Cell Lysates
  6. Diminished NO release in chronic hypoxic human endothelial cells.
    Authors: Ostergaard L, Stankevicius E, Andersen MR, Eskildsen-Helmond Y, Ledet T, Mulvany MJ, Simonsen U
    Am. J. Physiol. Heart Circ. Physiol., 2007-08-24;293(5):H2894-903.
    Species: Human
    Sample Types: Cell Lysates
  7. The new oral immunomodulating drug DiNAC induces brachial artery vasodilatation at rest and during hyperemia in hypercholesterolemic subjects, likely by a nitric oxide-dependent mechanism.
    Authors: Pettersson K, Kjerrulf M, Jungersten L, Johansson K, Langstrom G, Kalies I, Lenkei R, Walldius G, Lind L
    Atherosclerosis, 2006-12-08;196(1):275-82.
    Species: Human
    Sample Types: Cell Lysates
  8. High-density lipoprotein cholesterol regulates endothelial progenitor cells by increasing eNOS and preventing apoptosis.
    Authors: Noor R, Shuaib U, Wang CX, Todd K, Ghani U, Schwindt B, Shuaib A
    Atherosclerosis, 2006-08-01;192(1):92-9.
    Species: Human
    Sample Types: Cell Lysates
  9. Neuropeptide (calcitonin gene-related peptide) induction of nitric oxide in human keratinocytes in vitro.
    Authors: E Y, Golden SC, Shalita AR, Lee WL, Maes DH, Matsui MS
    J. Invest. Dermatol., 2006-05-18;126(9):1994-2001.
    Species: Human
    Sample Types: Cell Lysates
  10. Effective treatment of vascular endothelial growth factor refractory hindlimb ischemia by a mutant endothelial nitric oxide synthase gene.
    Authors: Qian HS, Liu P, Huw LY, Orme A, Halks-Miller M, Hill SM, Jin F, Kretschmer P, Blasko E, Cashion L, Szymanski P, Vergona R, Harkins R, Yu J, Sessa WC, Dole WP, Rubanyi GM, Kauser K
    Gene Ther., 2006-04-27;13(18):1342-50.
    Species: Rat
    Sample Types: Tissue Homogenates
  11. Relationship between body composition and skeletal muscle eNOS.
    Authors: Hickner RC, Kemeny G, Stallings HW, Manning SM, McIver KL
    Int J Obes (Lond), 2006-02-01;30(2):308-12.
    Species: Human
    Sample Types: Tissue Homogenates
  12. Effects of hyperthermia pretreatment on expression of heme oxygenase-1 and nitric oxide synthase in rats subjected to experimental anaphylactic shock.
    Authors: Yang RC, Chang CY, Lu TS, Chen SC
    Chin J Physiol, 2005-12-31;48(4):193-9.
    Species: Rat
    Sample Types: Tissue Homogenates
  13. Endothelial nitric oxide synthase content in adipose tissue from obese and lean African American and white American women.
    Authors: Galvin VB, Barakat H, Kemeny G, Macdonald KG, Pories WJ, Hickner RC
    Metab. Clin. Exp., 2005-10-01;54(10):1368-73.
    Species: Human
    Sample Types: Tissue Homogenates
  14. Dietary fish oil prevents vascular dysfunction and oxidative stress in hyperinsulinemic rats.
    Authors: Nyby MD, Matsumoto K, Yamamoto K, Abedi K, Eslami P, Hernandez G, Smutko V, Berger ME, Tuck ML
    Am. J. Hypertens., 2005-02-01;18(2):213-9.
    Species: Rat
    Sample Types: Tissue Homogenates
  15. 15-deoxy-delta12,14-prostaglandin-J2 inhibits expression of eNOS in human endothelial cells.
    Authors: Jozkowicz A, Nigisch A, Winter B, Weigel G, Hukb I, Dulaka J
    Prostaglandins Other Lipid Mediat., 2004-10-01;74(1):11-28.
    Species: Human
    Sample Types: Cell Lysates
  16. Reduced endothelial nitric oxide synthase activity and concentration in fetal umbilical veins from maternal cigarette smokers.
    Authors: Andersen MR, Walker LR, Stender S
    Am. J. Obstet. Gynecol., 2004-07-01;191(1):346-51.
    Species: Human
    Sample Types: Cell Lysates
  17. Inflammation and nitric oxide production in skeletal muscle of type 2 diabetic patients.
    Authors: Torres SH, De Sanctis JB, de L Briceno M, Hernandez N, Finol HJ
    J. Endocrinol., 2004-06-01;181(3):419-27.
    Species: Human
    Sample Types: Tissue Homogenates
  18. Bronchiolitis obliterans induced by intratracheal papaverine: a novel animal model.
    Authors: Svetlecic J, Molteni A, Herndon B
    Lung, 2004-01-01;182(2):119-34.
    Species: Rat
    Sample Types: Tissue Homogenates
  19. Interaction between nitric oxide and cyclooxygenase pathways in endothelial cells.
    Authors: L&amp;apos;Abbate A
    J. Vasc. Res., 2003-10-28;40(5):491-9.
    Species: Human
    Sample Types: Cell Lysates
  20. Oxidative injury due to chronic nitric oxide synthase inhibition in rat: effect of regular exercise on the heart.
    Authors: Husain K, Hazelrigg SR
    Biochim. Biophys. Acta, 2002-05-21;1587(1):75-82.
    Species: Rat
    Sample Types: Tissue Homogenates
  21. Retinal vascular endothelial growth factor induces intercellular adhesion molecule-1 and endothelial nitric oxide synthase expression and initiates early diabetic retinal leukocyte adhesion in vivo.
    Authors: Joussen AM, Poulaki V, Qin W, Kirchhof B, Mitsiades N, Wiegand SJ, Rudge J, Yancopoulos GD, Adamis AP
    Am. J. Pathol., 2002-02-01;160(2):501-9.
    Species: Rat
    Sample Types: Tissue Homogenates

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