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Mouse Fas Ligand/TNFSF6 Quantikine ELISA Kit

Catalog #: MFL00
13 Citations Datasheet / COA / SDS
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MFL00
Mouse Fas Ligand/TNFSF6 ELISA Standard Curve
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Citations (13)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Mouse Fas Ligand/TNFSF6 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 (50 uL), EDTA Plasma (50 uL)
Sensitivity
3.9 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
Specificity
Natural and recombinant mouse Fas Ligand
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 Mouse Fas Ligand Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse Fas Ligand in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse Fas Ligand and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse Fas Ligand{\f9\i 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 mouse Fas Ligand.

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 Culture Supernates, Serum, EDTA Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 84 261 675 76 243 663
Standard Deviation 8.1 9 20 6.8 12 33
CV% 9.6 3.4 3 8.9 4.9 5

Recovery

The recovery of mouse Fas Ligand spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 107 84-120
EDTA Plasma (n=6) 99 87-110
Serum (n=6) 98 86-111

Linearity

To assess the linearity of the assay, six samples containing and/or spiked with high concentrations of mouse Fas Ligand in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse Fas Ligand/TNFSF6 ELISA Linearity

Scientific Data

N/A Mouse Fas Ligand/TNFSF6 ELISA Standard Curve View Larger

Mouse Fas Ligand/TNFSF6 ELISA Standard Curve

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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 2 - 8 °C. Do not use past expiration date.

Background: Fas Ligand/TNFSF6

Fas Ligand (FasL), also known as CD178, CD95L, or TNFSF6, is a transmembrane protein homotrimer that binds to Fas/CD95 and triggers apoptosis in the Fas-expressing cell. Fas Ligand also binds the soluble decoy receptor DcR3. It is expressed on activated Th1 cells, CD8+ cytotoxic T cells, and NK cells. A soluble form can be released which remains trimeric and retains the ability to bind Fas. Fas Ligand-induced apoptosis plays a central role in the development of immune tolerance and the maintance of immune privileged sites. Tumor cells can evade immune surveillance by upregulating Fas Ligand to kill tumor infiltrating lymphocytes. In gld mice, a Fas Ligand point mutation is the cause of severe lymphoproliferation and systemic autoimmunity.

Entrez Gene IDs:
356 (Human); 14103 (Mouse); 25385 (Rat)
Alternate Names:
apoptosis (APO-1) antigen ligand 1; Apoptosis antigen ligand; APT1LG1CD95L; APTL; CD178 antigen; CD178; CD95L; CD95-L; Fas antigen ligand; Fas ligand (TNF superfamily, member 6); Fas Ligand; FASLCD95 ligand; FASLG; TNFSF6; TNFSF6FasL; tumor necrosis factor (ligand) superfamily, member 6; tumor necrosis factor ligand superfamily member 6
&#9888; 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.
  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 the appropriate 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 on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
    6. 100 µL Conjugate
  6.   Add 100 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 5 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   Add 100 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 Mouse Fas Ligand/TNFSF6 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.

13 Citations: Showing 1 - 10
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  1. Lung Injury Combined with Loss of Regulatory T Cells Leads to De Novo Lung-Restricted Autoimmunity
    J Immunol, 2016-05-18;0(0):.
    Species: Mouse
    Sample Types: BALF
  2. Hematopoietic Fas deficiency does not affect experimental atherosclerotic lesion formation despite inducing a proatherogenic state.
    Authors: de Claro RA, Zhu X, Tang J, Morgan-Stevenson V, Schwartz BR, Iwata A, Liles WC, Raines EW, Harlan JM
    Am. J. Pathol., 2011-05-06;178(6):2931-7.
    Species: Mouse
    Sample Types: Plasma
  3. TLR4 through IFN-beta promotes low molecular mass hyaluronan-induced neutrophil apoptosis.
    Authors: Leu SW, Shi L, Xu C, Zhao Y, Liu B, Li Y, Shiedlin A, Xiang C, Shen H, Quinn DA, Hales CA, Zhao H
    J. Immunol., 2010-11-22;186(1):556-62.
    Species: Mouse
    Sample Types: BALF
  4. Ectodomain shedding of EGFR ligands and TNFR1 dictates hepatocyte apoptosis during fulminant hepatitis in mice.
    Authors: Murthy A, Defamie V, Smookler DS
    J. Clin. Invest., 2010-07-12;120(8):2731-44.
    Species: Mouse
    Sample Types: Serum
  5. Critical roles of inflammation and apoptosis in improved survival in a model of hyperoxia-induced acute lung injury in Pneumocystis murina-infected mice.
    Authors: Beck JM, Preston AM, Wilcoxen SE, Morris SB, Sturrock A, Paine R
    Infect. Immun., 2009-01-05;77(3):1053-60.
    Species: Mouse
    Sample Types: Tissue Homogenates
  6. Mechanical ventilation enhances lung inflammation and caspase activity in a model of mouse pneumovirus infection.
    Authors: Bem RA, van Woensel JB, Bos AP, Koski A, Farnand AW, Domachowske JB, Rosenberg HF, Martin TR, Matute-Bello G
    Am. J. Physiol. Lung Cell Mol. Physiol., 2008-11-07;296(1):L46-56.
    Species: Mouse
    Sample Types: Tissue Homogenates
  7. Ozone inhalation induces exacerbation of eosinophilic airway inflammation and hyperresponsiveness in allergen-sensitized mice.
    Authors: Kierstein S, Krytska K, Sharma S, Amrani Y, Salmon M, Panettieri RA, Zangrilli J, Haczku A
    Allergy, 2008-04-01;63(4):438-46.
    Species: Mouse
    Sample Types: BALF
  8. Tissue inhibitor of metalloproteinases-3 facilitates Fas-mediated neuronal cell death following mild ischemia.
    Authors: Wetzel M, Li L, Harms KM, Roitbak T, Ventura PB, Rosenberg GA, Khokha R, Cunningham LA
    Cell Death Differ., 2007-10-26;15(1):143-51.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. The Fas/Fas ligand system inhibits differentiation of murine osteoblasts but has a limited role in osteoblast and osteoclast apoptosis.
    Authors: Kovacic N, Lukic IK, Grcevic D, Katavic V, Croucher P, Marusic A
    J. Immunol., 2007-03-15;178(6):3379-89.
    Species: Mouse
    Sample Types: Cell Lysates
  10. Targeted cardiac expression of soluble Fas prevents the development of heart failure in mice with cardiac-specific expression of MCP-1.
    Authors: Niu J, Azfer A, Deucher MF, Goldschmidt-Clermont PJ, Kolattukudy PE
    J. Mol. Cell. Cardiol., 2006-05-05;40(6):810-20.
    Species: Mouse
    Sample Types: Plasma
  11. Suppression of graft coronary artery disease by a brief treatment with a selective epsilonPKC activator and a deltaPKC inhibitor in murine cardiac allografts.
    Authors: Tanaka M, Terry RD, Mokhtari GK, Inagaki K, Koyanagi T, Kofidis T, Mochly-Rosen D, Robbins RC
    Circulation, 2004-09-14;110(11):II194-9.
    Species: Mouse
    Sample Types: Tissue Homogenates
  12. Cardiomyocyte-specific Bcl-2 overexpression attenuates ischemia-reperfusion injury, immune response during acute rejection, and graft coronary artery disease.
    Authors: Tanaka M, Nakae S, Terry RD, Mokhtari GK, Gunawan F, Balsam LB, Kaneda H, Kofidis T, Tsao PS, Robbins RC
    Blood, 2004-07-27;104(12):3789-96.
    Species: Mouse
    Sample Types: Tissue Homogenates
  13. T cell-specific ablation of Fas leads to Fas ligand-mediated lymphocyte depletion and inflammatory pulmonary fibrosis.
    Authors: Hao Z, Hampel B, Yagita H, Rajewsky K
    J. Exp. Med., 2004-05-17;199(10):1355-65.
    Species: Mouse
    Sample Types: Serum

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