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Fetuin A ELISA Kit

AHSG Reaktivität: Human Colorimetric Sandwich ELISA 5.0-370 ng/mL Cell Culture Supernatant, Plasma, Serum, Urine
Produktnummer ABIN1305166
  • Target Alle Fetuin A (AHSG) ELISA Kits anzeigen
    Fetuin A (AHSG) (alpha-2-HS-Glycoprotein (AHSG))
    Reaktivität
    • 8
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    5.0-370 ng/mL
    Untere Nachweisgrenze
    5 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    This ELISA (enzyme-linked immunosorbent assay) kit is intended for the quantitative determination of human Fetuin-A, also known as alpha-2-HS glycoprotein (AHSG), in serum, plasma, cell culture supernatant, tissue extraction and urine. The measurement of Fetuin-A in serum or plasma aids in the diagnosis of some cancers and genetically inherited deficiencies of this serum protein. This Fetuin-A ELISA kit is for laboratory professional use.
    Marke
    ED™
    Proben
    Serum, Plasma, Cell Culture Supernatant, Urine
    Analytische Methode
    Quantitative
    Produktmerkmale
    Cardiovascular Disease
    Bestandteile
    1. Fetuin-A Antibody Coated Microplate. One bottle contains 12 mL of 0.5 M sulfuric acid. This reagent should be stored at 2-8 °C or room temperature and is stable until the expiration date on the kit box
    Fetuin-A Standards.Five vials each containing 0.5 mL of human Fetuin-A in a liquid bovine serum based matrix with a non-azide preservative. Refer to vial for exact concentration for each standard. All the standards should be stored at -20 °C or below after the first use with up to 3 freeze cycles Fetuin-A Controls.Two vials each containing 0.5mL of human Fetuin-A in a liquid bovine serum based matrix with a non-azide preservative. Refer to vials for exact concentration range for each control. Both controls should be stored at -20 °C or below after the first use with up to 3 freeze cycles.
    Benötigtes Material
    1. Precision single channel pipettes capable of delivering 10 μL, 25 μL, 100 μL, and 1000 μL
    2. Repeating dispenser suitable for delivering 100 μL
    3. Disposable pipette tips suitable for above volume dispensing
    4. Disposable 12 x 75 mm or 13 x 100 glass or plastic tubes
    5. Disposable plastic 100 mL and 1000 mL bottle with caps
    6. Aluminum foil
    7. Deionized or distilled water
    8. Plastic microtiter well cover or polyethylene film
    9. ELISA multichannel wash bottle or automatic (semi-automatic) washing system
    10. Spectrophotometric microplate reader capable of reading absorbance at 450 nm
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  • Probenmenge
    10 μL
    Testdauer
    4 h
    Plattentyp
    Pre-coated
    Protokoll
    Assay standards, controls and prediluted patient serum samples containing human Fetuin-A are added to microtiter wells of microplate that was coated with a high affinity polyclonal goat anti-human Fetuin-A antibody. After the first incubation period, the antibody on the wall of microtiter well captures human Fetuin-A in the sample and unbound protein in each microtiter well is washed away. Then a horseradish peroxidase (HRP) conjugated polyclonal anti-human Fetuin-A antibody is added to each microtiter well and a sandwich of capture antibody - human Fetuin-A - HRP conjugated tracer antibody is formed. The unbound tracer antibody is removed in the subsequent washing step. HRP conjugated tracer antibody bound to the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the tracer antibody bound to the Fetuin-A on the wall of the microtiter well is directly proportional to the amount of Fetuin-A in the sample. A standard curve is generated by plotting the absorbance versus the respective human Fetuin-A concentration for each standard on point-to-point or cubical scales. The concentration of human Fetuin-A in test samples is determined directly from this standard curve.
    Aufbereitung der Reagenzien

    (1) Prior to use allow all reagents to come to room temperature. Reagents from different kit lot numbers should not be combined or interchanged.
    (2) Fetuin-A Assay Buffer Concentrate and ELISA Wash Concentrate must be diluted to working solution prior to use. Please see REAGENTS section for details

    Probennahme
    Only 10 μL of human serum or plasma is required for human Fetuin-A measurement. No special preparation of individual is necessary prior to specimen collection. Whole blood should be collected and must be allowed to clot for minimum 30 minutes at room temperature before the serum is separated by centrifugation (850 1500xg for 10 minutes). The serum should be separated from the clot within three hours of blood collection and transferred to a clean test tube. Serum samples may be stored at 20 °C or below until measurement. Avoid more than three freeze-thaw cycles of specimen. Twenty four hour urine sample is recommended to be used for the determination of urine Fetuin-A concentration. Spot urine from the second morning urination may be used if strenuous physical activity shortly before sample collection has been ruled out and polyuric renal dysfunction is not present. Intra-individual day-to-day fluctuations in the concentration of urine proteins caused by diurisis may be reduced by relating to the urinary creatinine concentration.For cell culture supernatant, tissue extracts, one should serial dilute test sample and measure multiple diluted samples for a more accurate Fetuin-A test result.
    Aufbereitung der Proben

    Patient serum or plasma sample needs to be diluted 1:10,000 with assay buffer before being measured.
    (1) Label 2 test tubes (12x75 mm) with 1A and 1B.
    (2) Add 1 mL of assay buffer to each tube (both 1A and 1B).
    (3) Pipet 10 μL of patient serum or plasma sample to tube 1A and mix well (1:100 dilution).(4) Pipet 10 μL of diluted patient sample from 1A to tube 1B mix well (1:10,000 dilution).Note: It is recommended to use a precision/calibrated pipette and careful technique to perform the dilution in order to get precise results! We recommend using Eppendorf Repeat Pipette with 12.5 mL combitip for adding 1 mLassay buffer. 50 mL combitip is not recommended. Patient urine sample needs to be diluted 1:100 with assay buffer before being measured.
    (1) Label 1 test tube (12x75 mm) with 1.
    (2) Add 1 mL of assay buffer to tube.
    (3) Pipet 10 μL of patient urine sample to tube 1 and mix well (1:100 dilution).Note: If a higher than standard level 5 of Fetuin-A test result is obtained, a further dilution of urine sample (e.g. 1:500) should be measured for reporting a more accurate test result.

    Testdurchführung

    (1) Place a sufficient number of antibody coated microwell strips in a holder to run human Fetuin-A standards, controls and unknown samples in duplicate.
    (2) Test Configuration
    (3) Add 25 μL of standards (Cat. 30001-30005), controls (Cat. 30007-30008) and 1:10,000 diluted patient samples into the designated microwell. Note: if urine sample is used, 1:100 diluted urine sample should be used.
    (4) Add 100 μL of assay buffer to each well.
    (5) Mix gently and cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
    (6) Incubate the plate at room temperature for 2 hours.
    (7) Prepare Tracer Antibody Working Solution by 1:21 fold dilution of the Fetuin-A Tracer Antibody with the Tracer Antibody Diluent. For each strip, it is required to mix 1 mL of Tracer Antibody Diluent with 50 μL of Fetuin-A Tracer Antibody in a clean test tube.
    (8) Remove the aluminum foil and the plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 μL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
    (9) Add 100 μL of above diluted tracer antibody working solution to each of the wells.
    (10) Cover the plate with one plate sealer and also with aluminum foil to avoid exposure to light.
    (11) Incubate the plate at room temperature for 30 minutes.
    (12) Remove the aluminum foil and the plate sealer. Aspirate the contents of each well. Wash each well 5 times by dispensing 350 μL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
    (13) Add 100 μL of ELISA HRP Substrate into each of the wells.
    (14) Cover the plate with aluminum foil to avoid exposure to light.
    (15) Incubate the plate at room temperature for 20 minutes
    (16) Remove the aluminum foil. Add 100 μL of ELISA Stop Solution into each of the wells. Mix gently.
    (17) Read the absorbance at 450 nm within 10 minutes in a microplate readerNOTE: to reduce the background, one can set the instrument to dual wavelength measurement at 450 nm with background wavelength correction set at 595 nm, 620 nm or 630 nm.

    Ergebnisberechnung
    1. Calculate the average absorbance for each pair of duplicate test results
      2. Subtract the average absorbance of the STD 1 (0 ng/mL) from the average absorbance of all other readings to obtain corrected absorbance.
      3. The standard curve is generated by the corrected absorbance of all standard levels on the ordinate against the standard concentration on the abscissa using point-to-point or log-log paper. Appropriate computer assisted data reduction programs may also be used for the calculation of results.
      4. We recommend a linear ordinate for optical density and a linear abscissa for concentration. The human serum or plasma Fetuin-A concentrations for the controls and 1:10,000 diluted samples are read directly from the standard curve using their respective corrected absorbance.
    Testpräzision
    The intra-assay precision is validated by measuring two samples in a single assay with 20 replicate determinations. The inter-assay precision is validated by measuring two samples in duplicate in 12 individual assays.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Vorsichtsmaßnahmen
    The reagents must be used in a clinical reference laboratory and are intended for in vitro diagnostic use by medical or laboratory professionals only. The source material for reagents containing bovine serum was derived in the contiguous 48 United States. It was obtained only from healthy donor animals maintained under veterinary supervision and found free of contagious diseases. Wear gloves while performing this assay and handle these reagents as if they are potentially infectious. Avoid contact with reagents containing TMB, hydrogen peroxide, or sulfuric acid. TMB may cause irritation to skin and mucous membranes and cause an allergic skin reaction. TMB is a suspected carcinogen. Sulfuric acid may cause severe irritation on contact with skin. Do not get in eyes, on skin, or on clothing. Do not ingest or inhale fumes. On contact, flush with copious amounts of water for at least 15 minutes. Use Good Laboratory Practices.
    Lagerung
    4 °C
  • Schoppet, Rauner, Benner, Chapurlat, Hofbauer, Szulc: "Serum fetuin-A levels and abdominal aortic calcification in healthy men - The STRAMBO study." in: Bone, Vol. 79, pp. 196-202, (2015) (PubMed).

    Sonmez, Nikolic, Dogru, Ercin, Genc, Cesur, Tapan, Karslioğlu, Montalto, Banach, Toth, Bagci, Rizzo: "Low- and high-density lipoprotein subclasses in subjects with nonalcoholic fatty liver disease." in: Journal of clinical lipidology, Vol. 9, Issue 4, pp. 576-82, (2015) (PubMed).

    Liu, Lamendola, Ariel, Abbasi, Kim, Cardell, Tomasso, Xu, Patel, Mojaddidi, Grove, Kushida, Reaven: "Usefulness of fetuin-A to predict risk for cardiovascular disease among patients with obstructive sleep apnea." in: The American journal of cardiology, Vol. 116, Issue 2, pp. 219-24, (2015) (PubMed).

    Celebi, Genc, Gurel, Sertoglu, Kara, Tapan, Acikel, Karslioglu, Ercin, Dogru: "The relationship of circulating fetuin-a with liver histology and biomarkers of systemic inflammation in nondiabetic subjects with nonalcoholic fatty liver disease." in: Saudi journal of gastroenterology : official journal of the Saudi Gastroenterology Association, Vol. 21, Issue 3, pp. 139-45, (2015) (PubMed).

    Fink, Bůžková, Garimella, Mukamal, Cauley, Kizer, Barzilay, Jalal, Ix: "Association of Fetuin-A With Incident Fractures in Community-Dwelling Older Adults: The Cardiovascular Health Study." in: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 30, Issue 8, pp. 1394-402, (2015) (PubMed).

    Kurnatowska, Grzelak, Masajtis-Zagajewska, Kaczmarska, Stefa?czyk, Vermeer, Maresz, Nowicki: "Effect of vitamin K2 on progression of atherosclerosis and vascular calcification in nondialyzed patients with chronic kidney disease stages 3-5." in: Polskie Archiwum Medycyny Wewne?trznej, Vol. 125, Issue 9, pp. 631-40, (2015) (PubMed).

    Cuthbertson, Shojaee-Moradie, Sprung, Jones, Pugh, Richardson, Kemp, Barrett, Jackson, Thomas, Bell, Umpleby: "Dissociation between exercise-induced reduction in liver fat and changes in hepatic and peripheral glucose homeostasis in obese patients with Non-Alcoholic Fatty Liver Disease." in: Clinical science (London, England : 1979), (2015) (PubMed).

    Laugsand, Ix, Bartz, Djousse, Kizer, Tracy, Dehghan, Rexrode, Lopez, Rimm, Siscovick, ODonnell, Newman, Mukamal, Jensen: "Fetuin-A and risk of coronary heart disease: A Mendelian randomization analysis and a pooled analysis of AHSG genetic variants in 7 prospective studies." in: Atherosclerosis, Vol. 243, Issue 1, pp. 44-52, (2015) (PubMed).

    Debowska, Poleszczuk, Wojcik-Zaluska, Ksiazek, Zaluska: "Phosphate Kinetics During Weekly Cycle of Hemodialysis Sessions: Application of Mathematical Modeling." in: Artificial organs, (2015) (PubMed).

    Barros, Dirrichs, Koos, Reinartz, Kaesler, Kramann, Gladziwa, Ketteler, Floege, Marx, Torregrosa, Keszei, Brandenburg: "Epicardial adipose tissue in long-term hemodialysis patients: its association with vascular calcification and long-term development." in: Journal of nephrology, (2015) (PubMed).

    Scialla, Kao, Crainiceanu, Sozio, Oberai, Shafi, Coresh, Powe, Plantinga, Jaar, Parekh: "Biomarkers of vascular calcification and mortality in patients with ESRD." in: Clinical journal of the American Society of Nephrology : CJASN, Vol. 9, Issue 4, pp. 745-55, (2014) (PubMed).

    Laughlin, McEvoy, Barrett-Connor, Daniels, Ix: "Fetuin-A, a new vascular biomarker of cognitive decline in older adults." in: Clinical endocrinology, Vol. 81, Issue 1, pp. 134-40, (2014) (PubMed).

    Mutlu, Vuralkan, Yardim Akaydin, Akin, Miser: "Effects of adenoid/tonsillectomy on inflammatory response in snoring children with witnessed apnoea." in: Clinical otolaryngology : official journal of ENT-UK ; official journal of Netherlands Society for Oto-Rhino-Laryngology & Cervico-Facial Surgery, Vol. 39, Issue 5, pp. 266-71, (2014) (PubMed).

    Ix, Biggs, Mukamal, Kizer, Zieman, Siscovick, Mozzaffarian, Jensen, Nelson, Ruderman, Djousse: "Association of fetuin-a with incident diabetes mellitus in community-living older adults: the cardiovascular health study." in: Circulation, Vol. 125, Issue 19, pp. 2316-22, (2012) (PubMed).

    Ix, Katz, de Boer, Kestenbaum, Peralta, Jenny, Budoff, Allison, Criqui, Siscovick, Shlipak: "Fetuin-A is inversely associated with coronary artery calcification in community-living persons: the Multi-Ethnic Study of Atherosclerosis." in: Clinical chemistry, Vol. 58, Issue 5, pp. 887-95, (2012) (PubMed).

  • Target Alle Fetuin A (AHSG) ELISA Kits anzeigen
    Fetuin A (AHSG) (alpha-2-HS-Glycoprotein (AHSG))
    Andere Bezeichnung
    Fetuin-A (AHSG Produkte)
    Synonyme
    ahs ELISA Kit, a2hs ELISA Kit, hsga ELISA Kit, fetua ELISA Kit, wu:fb63g02 ELISA Kit, zgc:103687 ELISA Kit, AHSG ELISA Kit, MGC116429 ELISA Kit, A2HS ELISA Kit, AHS ELISA Kit, FETUA ELISA Kit, HSGA ELISA Kit, Aa2-066 ELISA Kit, pp63 ELISA Kit, alpha-2-HS-glycoprotein ELISA Kit, alpha 2-HS glycoprotein ELISA Kit, alpha-2-HS-glycoprotein 1 ELISA Kit, alpha-2-HS-glycoprotein L homeolog ELISA Kit, Cphamn1_1981 ELISA Kit, AHSG ELISA Kit, ahsg ELISA Kit, ahsg1 ELISA Kit, ahsg.L ELISA Kit, Ahsg ELISA Kit
    Hintergrund
    Fetuin-A, also known as alpha-2-HS glycoprotein, is a 59 kDa glycoprotein that consists of two amino-terminal cystatin domains and a smaller carboxyl-terminal domain. Fetuin-A is synthesized by the liver and secreted into blood stream, where its concentration in adult mammals ranges from 0.5 1.0 g/L. Fetuin-A occurs in high serum concentration during fetal life and involves in protease inhibitory activities and development-associated regulation of calcium metabolism and osteogenesis. It accumulates in bones and teeth as a major fraction of noncollagenous bone proteins. Biologically, studies have demonstrated that Fetuin-A is the major calcification inhibitor found in circulation, where it interferes with calcium salt precipitation. Recent study has indicated that Fetuin-A level drops in uremic patients on hemodialysis in comparison to normal healthy controls. The low Fetuin-A level in patients with chronic kidney failure strongly associates with a higher cardiovascular mortality. On the other hand, it is demonstrated that a higher than normal serum Fetuin-A in older population is associated with diabetes, which is independent from other markers of insulin resistance. Further, a higher Fetuin-A level may be an independent risk marker of patients with cardiovascular diseases.
    Molekulargewicht
    59 kDa
    Gen-ID
    197
    NCBI Accession
    NP_001613.2
    UniProt
    P02765
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