Human Fas/TNFRSF6/CD95 Quantikine ELISA Kit

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Human Fas/TNFRSF6/CD95 ELISA Standard Curve
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Human Fas/TNFRSF6/CD95 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 (100 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL)
Sensitivity
20 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human sFas
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 sFas Immunoassay is a 4.5 hour solid phase ELISA designed to measure human sFas in cell culture supernates, serum, and plasma. It contains recombinant human Fas/Fc chimera expressed from NS0 cells and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained measuring natural human sFas showed dose-response curves that were parallel to the standard curves obtained using the recombinant Fas/Fc chimera. These results indicate that this kit can be used to determine relative mass values for natural human sFas.

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, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 130 804 1424 140 811 1391
Standard Deviation 5 23.3 65.2 9.4 31.8 40
CV% 3.8 2.9 4.6 6.7 3.9 2.9

Recovery

The recovery of Fas/Fc chimera spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 92-107
Citrate Plasma (n=5) 98 82-116
EDTA Plasma (n=5) 97 86-111
Heparin Plasma (n=5) 94 88-109
Serum (n=5) 97 91-110

Linearity

To assess the linearity of the assay, five samples containing or spiked with high concentrations of sFas or Fas/Fc chimera were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Fas/TNFRSF6/CD95 ELISA Linearity

Data Examples

Human Fas/TNFRSF6/CD95 ELISA Standard Curve

Product Datasheets

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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/TNFRSF6/CD95

Fas (Fibroblast-associated), also known as Apo-1, CD95, and TNFRSF6, was originally identified as a cell-surface protein that binds to monoclonal antibodies that are cytolytic for various human cell lines. Alternatively spliced cDNAs encoding multiple Fas isoforms, including a soluble form of Fas, have been identified. Fas is highly expressed in epithelial cells, hepatocyes, activated mature lymphocytes, virus-transformed lymphocytes and other tumor cells. Fas expression has also been detected in mouse thymus, liver, heart, lung, kidney and ovary.

Long Name:
Fibroblast-associated
Entrez Gene IDs:
355 (Human); 14102 (Mouse); 246097 (Rat); 102140989 (Cynomolgus Monkey); 493881 (Feline)
Alternate Names:
Apo-1 antigen; Apo-1; apoptosis antigen 1; Apoptosis-mediating surface antigen FAS; APT1; APT1FASTM; CD95 antigen; CD95; CD95ALPS1A; Fas (TNF receptor superfamily, member 6); Fas AMA; Fas antigen; Fas; FAS1; FASLG receptor; TNFRSF6; TNFRSF6member 6; tumor necrosis factor receptor superfamily member 6
⚠ 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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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 Fas/TNFRSF6/CD95 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.

17 Citations: Showing 1 - 10
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  1. Tumor necrosis factor-alpha and Fas/Fas ligand signaling pathways in chronic spontaneous urticaria
    Authors: R Grzanka, A Damasiewic, A Kasperska-
    Allergy Asthma Clin Immunol, 2019;15(0):15.
    Species: Human
    Sample Types: Serum
  2. Evaluation of soluble concentration Fas and Fas ligand in maternal and cord blood 3rd trimester of pregnancy
    Authors: A Karowicz-B, U Kowalska-K, D Estemberg, A Sikora-Szu
    Ginekol. Pol., 2018;89(3):142-146.
    Species: Human
    Sample Types: Serum
  3. FAS-antisense 1 lncRNA and production of soluble versus membrane Fas in B-cell lymphoma.
    Authors: Sehgal, L, Mathur, R, Braun, F K, Wise, J F, Berkova, Z, Neelapu, S, Kwak, L W, Samaniego, F
    Leukemia, 2014;28(12):2376-87.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. High admission glucose levels increase Fas apoptosis and mortality in patients with acute ST-elevation myocardial infarction: a prospective cohort study.
    Authors: Chang J, Zhang G, Zhang L, Hou Y, Liu X, Zhang L
    Cardiovasc Diabetol, 2013;12(1):171.
    Species: Human
    Sample Types: Plasma
  5. Effect of quinapril on in-stent restenosis and relation to plasma apoptosis signaling molecules.
    Authors: Deftereos S, Giannopoulos G, Kossyvakis C, Kaoukis A, Raisakis K, Driva M, Panagopoulou V, Lappos S, Rentoukas I, Pyrgakis V, Alpert MA
    Am. J. Cardiol., 2010;105(1):54-8.
    Species: Human
    Sample Types: Plasma
  6. Soluble and membrane levels of molecules involved in the interaction between clonal plasma cells and the immunological microenvironment in multiple myeloma and their association with the characteristics of the disease.
    Authors: Perez-Andres M, Almeida J, Martin-Ayuso M, De Las Heras N, Moro MJ, Martin-Nunez G, Galende J, Cuello R, Abuin I, Moreno I, Dominguez M, Hernandez J, Mateo G, San Miguel JF, Orfao A
    Int. J. Cancer, 2009;124(2):367-75.
    Species: Human
    Sample Types: Plasma
  7. Fas expression in conjunctival epithelial cells of patients with cystic fibrosis.
    J. Interferon Cytokine Res., 2009;29(11):735-40.
    Species: Human
    Sample Types: Serum
  8. Prognostic value of apoptosis markers in advanced heart failure patients.
    Authors: Niessner A, Hohensinner PJ, Rychli K, Neuhold S, Zorn G, Richter B, Hulsmann M, Berger R, Mortl D, Huber K, Wojta J, Pacher R
    Eur. Heart J., 2009;30(7):789-96.
    Species: Human
    Sample Types: Plasma
  9. A 60-s postconditioning protocol by percutaneous coronary intervention inhibits myocardial apoptosis in patients with acute myocardial infarction.
    Authors: Zhao WS, Xu L, Wang LF, Zhang L, Zhang ZY, Liu Y, Liu XL, Yang XC, Cui L, Zhang L
    Apoptosis, 2009;14(10):1204-11.
    Species: Human
    Sample Types: Plasma
  10. Effects of darbepoetin-alpha on plasma pro-inflammatory cytokines, anti-inflammatory cytokine interleukin-10 and soluble Fas/Fas ligand system in anemic patients with chronic heart failure.
    Authors: Kourea K, Parissis JT, Farmakis D, Panou F, Paraskevaidis I, Venetsanou K, Filippatos G, Kremastinos DT
    Atherosclerosis, 2007;199(1):215-21.
    Species: Human
    Sample Types: Plasma
  11. Soluble Fas--a promising novel urinary marker for the detection of recurrent superficial bladder cancer.
    Authors: Svatek RS, Herman MP, Lotan Y, Casella R, Hsieh JT, Sagalowsky AI, Shariat SF
    Cancer, 2006;106(8):1701-7.
    Species: Human
    Sample Types: Urine
  12. Growth hormone-induced reduction of soluble apoptosis mediators is associated with reverse cardiac remodelling and improvement of exercise capacity in patients with idiopathic dilated cardiomyopathy.
    Authors: Parissis JT, Adamopoulos S, Karatzas D, Paraskevaidis J, Livanis E, Kremastinos D
    Eur J Cardiovasc Prev Rehabil, 2005;12(2):164-8.
    Species: Human
    Sample Types: Plasma
  13. High-dose glucose-insulin-potassium treatment reduces myocardial apoptosis in patients with acute myocardial infarction.
    Authors: Zhang L, Li YH, Zhang HY, Chen ML, Gao MM, Hu AH, Yang HS
    Eur. J. Clin. Invest., 2005;35(3):164-70.
    Species: Human
    Sample Types: Plasma
  14. Level of sFas/APO 1 in serum and cerebrospinal fluid in multiple sclerosis.
    Authors: Mahovic D, Petravic D, Petelin Z, Zurak N, Horvat G, Hajnsek S
    Clin Neurol Neurosurg, 2004;106(3):230-2.
    Species: Human
    Sample Types: Serum
  15. Nitric oxide is associated with poor embryo quality and pregnancy outcome in in vitro fertilization cycles.
    Authors: Lee TH, Wu MY, Chen MJ, Chao KH, Ho HN, Yang YS
    Fertil. Steril., 2004;82(1):126-31.
    Species: Human
    Sample Types: Follicular Fluid
  16. Escape from cardiomyocyte apoptosis by enterovirus persistence due to elevated soluble Fas-receptors.
    Authors: Alter P, Maisch B
    Z Kardiol, 2004;93(7):524-32.
    Species: Human
    Sample Types: Serum
  17. Evaluation of caspase 1 and sFas serum levels in patients with systemic sclerosis: correlation with lung dysfunction, joint and bone involvement.
    Authors: Dziankowska-Bartkowiak B, Waszczykowska E, Zalewska A, Sysa-Jedrzejowska A
    Mediators Inflamm., 2003;12(6):339-43.
    Species: Human
    Sample Types: Serum

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Human Fas/TNFRSF6/CD95 Quantikine ELISA Kit
By Anonymous on 11/22/2021
Sample Tested: human floxp embryonal stem cell culture medium