Adiponectin (ADIPOQ) ELISA Kit

Details zu Produkt Nr. ABIN612653, Anbieter: Anmelden zum Anzeigen
Antigen
  • ACDC
  • ACRP30
  • ADIPQTL1
  • ADPN
  • APM-1
  • APM1
  • GBP28
  • apM-1
  • 30kDa
  • APN
  • Acdc
  • Acrp30
  • Ad
  • adipo
  • apM1
  • ADN
  • acrpl
  • adipo-b
  • adipoql
  • zgc:153584
  • adipoq
  • adiponectin, C1Q and collagen domain containing
  • adiponectin, C1Q and collagen domain containing, b
  • adiponectin, C1Q and collagen domain containing L homeolog
  • ADIPOQ
  • Adipoq
  • adipoqb
  • adipoq.L
Reaktivität
Human
Alternativen
Kits mit alternativen Reaktivitäten:
55
42
33
12
10
8
8
7
6
3
2
2
1
1
1
1
Methodentyp
Sandwich ELISA
Untere Nachweisgrenze
0.5 ng/mL
Applikation
ELISA
Optionen
Hersteller
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Hersteller Produkt- Nr.
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Verwendungszweck The AssayMax Human Adiponectin ELISA Kit is designed for detection of adiponectin in human urine, plasma, serum and cell culture supernatants
Marke AssayMax
Proben Plasma, Cell Culture Supernatant
Analytische Methode Quantitative
Nachweismethode Colorimetric
Spezifität This assay recognizes both natural and recombinant human adiponectin. It can detect both globular domain and full-length adiponectin.
Bestandteile Adiponectin Microplate: A 96-well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody against human adiponectin. Sealing Tapes: Each kit contains 3 pre-cut, pressure-sensitive sealing tapes, which can be cut to fit the format of the individual assay. Adiponectin Standard: Human adiponectin in a buffered protein base (800 ng, lyophilized). Biotinylated Adiponectin Antibody (100x): A 100-fold concentrated biotinylated polyclonal antibody against adiponectin (80µl). MIx Diluent Concentrate (10x): A 10-fold concentrated buffered protein base (30 ml). Wash Buffer Concentrate (20x): A 20-fold concentrated buffered surfactant (30 ml, 2 botlles). Streptavidin-Peroxidase Conjugate (SP Conjugate): A 100-fold concentrate (80µl). 1 Chromogen Substrate: A ready-to-use stabilized peroxidase chromogen substrate tetramethylbenzidine (8 ml). Stop Solution: A 0.5 N hydrochloric acid to stop the chromogen substrate reaction (12 ml).
Benötigtes Material Microplate reader capable of measuring absorbance at 450 nm. Pipettes (1-20 µL, 20-200 µL, 200-1000µLand multiple channel). Deionized or distilled reagent grade water.
Antigen
Andere Bezeichnung Adiponectin (ADIPOQ ELISA Kit Abstract)
Hintergrund Adiponectin, also known as Adipocyte Complement-Related Protein of 30kDa (ACRP30), is a secreted serum protein expressed exclusively in differentiated adipocytes. Studies indicated that decreased plasma adiponectin concentration is associated with obesity, insulin resistance , essential hypertension , inflammation and atherosclerosis , and acute myocardial infarction. On the other hand, increased adiponectin level leads to nephrotic syndrome.
Probenmenge 50 μL
Testdauer < 4 h
Plattentyp Pre-coated
Protokoll This assay employs a quantitative sandwich enzyme immunoassay technique that measures adiponectin in less than 4 hours. A polyclonal antibody specific for adiponectin has been pre-coated onto a microplate. Adiponectin in standards and samples is sandwiched by the immobilized antibody and biotinylated polyclonal antibody specific for adiponectin, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.
Aufbereitung der Reagenzien

Freshly dilute all reagents and bring all reagents to room temperature before use. If crystals have formed in the concentrate, mix gently until the crystals have completely dissolved. MIx Diluent Concentrate (10x): Dilute the MIx Diluent Concentrate 1:10 with reagent grade water. Store for up to 1 month at 2-8°C. Adiponectin Standard: Reconstitute the 800 ng of human adiponectin Standard with 4 ml of MIx Diluent to generate a standard stock of 200 ng/ml. Allow the standard to sit for 10 minutes with gentle agitation prior to making dilutions. Prepare duplicate or triplicate standard points by diluting stock (200 ng/ml) 1: 4 to produce a standard solution of 50 ng/ml. Prepare duplicate or triplicate standard points by serially diluting the Standard solution (50 ng/ml) 1:2 with equal volume of MIx Diluent to produce 25, 12.5, 6.25, 3.125, 1.56 and 0.78 ng/ml. MIx Diluent serves as the zero standard (0 ng/ml). Any remaining solution should be frozen at -20°C. Standard Point Dilution [Adiponectin] (ng/ml) P1 1 part Stock (200 ng/ml) + 3 parts MIx Diluent 50.00 P2 1 part P1 + 1 part MIx Diluent 25.00 P3 1 part P2 + 1 part MIx Diluent 12.50 P4 1 part P3 + 1 part MIx Diluent 6.25 P5 1 part P4 + 1 part MIx Diluent 3.13 P6 1 part P5 + 1 part MIx Diluent 1.56 P7 1 part P6 + 1 part MIx Diluent 0.78 P8 MIx Diluent 0.00 Biotinylated Adiponectin Antibody (100x): Spin down the biotinylated antibody briefly and dilute the desired amount of the antibody 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C. Wash Buffer Concentrate (20x): Dilute the Wash Buffer Concentrate 1:20 with reagent grade water. SP Conjugate (100x): Spin down the SP Conjugate briefly and dilute the desired amount of the conjugate 1:100 with MIx Diluent. Any remaining solution should be frozen at -20°C.

Probennahme Plasma: Collect plasma using one-tenth volume of 0.1 M sodium citrate as an anticoagulant. Centrifuge samples at 2000 x g for 10 minutes and assay. Dilute samples 1:500 into MIx Diluent. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. (EDTA or Heparin can also be used as anticoagulant.) Serum: Samples should be collected into a serum separator tube. After clot formation, centrifuge samples at 2000 x g for 10 minutes. Remove serum and assay. Dilute samples 1:500 into MIx Diluent. The undiluted samples can be stored at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles. Cell Culture Supernatants: Centrifuge cell culture media at 2000 x g for 10 minutes to remove debris. Collect supernatants and assay. Dilute samples 1:10 into MIx Diluent. Store samples at -20°C or below. Avoid repeated freeze-thaw cycles. Urine: Collect urine using sample pot. Centrifuge samples at 600 x g for 10 minutes and assay. Store samples at -20°C or below for up to 3 months. Avoid repeated freeze-thaw cycles.
Testdurchführung

Prepare all reagents, working standards and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature (20 - 30 °C). Remove excess microplate strips from the plate frame and return them immediately to the foil pouch with desiccant inside. Reseal the pouch securely to minimize exposure to water vapor and store in a vacuum desiccator. Add 50 µL of Standard or sample per well. Cover wells with a sealing tape and incubate for one hour. Start the timer after the last sample addition. Wash five times with 200 µL of Wash Buffer manually. Invert the plate each time and decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. If using a machine wash six times with 300 µL of Wash Buffer and then invert the plate, decant the contents, hit it 4-5 times on absorbent paper towel to completely remove the liquid. Add 50 µL of Biotinylated Adiponectin Antibody to each well and incubate for one hour. Wash a microplate as described above. Add 50 µL of Streptavidin-Peroxidase Conjugate per well and incubate for 30 minutes. Turn on the microplate reader and set up the program in advance. Wash a microplate as described above. Add 50 µL of Chromogen Substrate per well and incubate for about 10 minutes or till the optimal blue color density develops. Gently tap plate to ensure thorough mixing and break the bubbles in the well with pipette tip. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. Read the absorbance on a microplate reader at a wavelength of 450 nm immediately. If wavelength correction is available, subtract readings at 570 nm from those at 450 nm to correct optical imperfections. Otherwise, read the plate at 450 nm only. Please note that some unstable black particles may be generated at high concentration points after stopping the reaction for about 10 minutes, which will reduce the readings. 3

Ergebnisberechnung

Calculate the mean value of the triplicate for each standard and sample. To generate a Standard Curve from the initial reaction time, plot the graph using the standard concentrations on the x-axis and the corresponding mean 405 nm absorbance or change in 3 absorbance per minute (A/min) on the y-axis. The best-fit line can be determined by regression analysis of the linear portion of the curve. Determine the unknown sample concentration from the Standard Curve and multiply the value by the dilution factor of 5. Standard Curve The curve is provided for illustration only. A standard curve should be generated each time the assay is performed.

Testpräzision Intra-assay and inter-assay coefficients of variation were 4.2 % and 7.3 % respectively.
Beschränkungen Nur für Forschungszwecke einsetzbar
Handhabung The kit should not be used beyond the expiration date.
Lagerung 4 °C/-20 °C
Informationen zur Lagerung Store kit at 2-8°C or -20°C upon arrival up to the expiration date. Opened MIx Diluent may be stored for up to 1 month at 2-8°C. Store reconstituted reagents at -20°C or below. Opened unused strip wells may return to the foil pouch with the desiccant pack, reseal along zip-seal. May be stored for up to 1 month in a vacuum desiccator.
Bilder des Herstellers
ELISA image for Adiponectin (ADIPOQ) ELISA Kit (ABIN612653) Adiponectin (ADIPOQ) ELISA Kit
Produkt verwendet in: Çerman, Aktaş, Altunay, Arıcı, Tulunay, Ozturk: "Dietary glycemic factors, insulin resistance, and adiponectin levels in acne vulgaris." in: Journal of the American Academy of Dermatology, 2016 (PubMed).

El-Haggar, Farrag, Kotkata: "Effect of ketotifen in obese patients with type 2 diabetes mellitus." in: Journal of diabetes and its complications, Vol. 29, Issue 3, pp. 427-32, 2015 (PubMed).

Ou, Lu, Wu, Hung, Wu, Yang, Chang: "Both diabetes and fetuin-A are independently associated with increased risk of arterial stiffness." in: Clinica chimica acta; international journal of clinical chemistry, Vol. 445, pp. 133-8, 2015 (PubMed).

Li, Chang, Huang, Wu, Yang, Chao: "Tomato juice supplementation in young women reduces inflammatory adipokine levels independently of body fat reduction." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 31, Issue 5, pp. 691-6, 2015 (PubMed).

Ergin, Altan, Pilanci, Sirekbasan, Cortuk, Cizmecigil, Ersin, Elbey, Dinc, Habip, Turan, Arinci, Richt, Goossens, Karakullukcu, Kocak, Saribas, Koksal, Yilmaz, Kocazeybek: "The role of adenovirus 36 as a risk factor in obesity: the first clinical study made in the fatty tissues of adults in Turkey." in: Microbial pathogenesis, Vol. 80, pp. 57-62, 2015 (PubMed).

Fei, Ling, Hua, Ren: "Effects of soybean oligosaccharides on antioxidant enzyme activities and insulin resistance in pregnant women with gestational diabetes mellitus." in: Food chemistry, Vol. 158, pp. 429-32, 2014 (PubMed).

El-Haggar, Mostafa: "Adipokines and biochemical changes in Egyptian obese subjects: possible variation with sex and degree of obesity." in: Endocrine, 2014 (PubMed).

Konekov, Korolev, Shevchenko, Lapsina, Koroleva, Zonova, Prokofev: "[Cytokine gene polymorphism in type 2 diabetes mellitus in Russian women from eastern Europe]." in: Terapevticheskiĭ arkhiv, Vol. 84, Issue 10, pp. 14-22, 2012 (PubMed).

Tsao, Lodish, Fruebis: "ACRP30, a new hormone controlling fat and glucose metabolism." in: European journal of pharmacology, Vol. 440, Issue 2-3, pp. 213-21, 2002 (PubMed).

Allgemeine Veröffentlichungen Adamczak, Wiecek, Funahashi, Chudek, Kokot, Matsuzawa: "Decreased plasma adiponectin concentration in patients with essential hypertension." in: American journal of hypertension, Vol. 16, Issue 1, pp. 72-5, 2003 (PubMed).

Matsubara, Namioka, Katayose: "Decreased plasma adiponectin concentrations in women with low-grade C-reactive protein elevation." in: European journal of endocrinology / European Federation of Endocrine Societies, Vol. 148, Issue 6, pp. 657-62, 2003 (PubMed).

Kojima, Funahashi, Sakamoto, Miyamoto, Soejima, Hokamaki, Kajiwara, Sugiyama, Yoshimura, Fujimoto, Miyao, Suefuji, Kitagawa, Ouchi, Kihara, Matsuzawa, Ogawa: "The variation of plasma concentrations of a novel, adipocyte derived protein, adiponectin, in patients with acute myocardial infarction." in: Heart (British Cardiac Society), Vol. 89, Issue 6, pp. 667, 2003 (PubMed).

Zoccali, Mallamaci, Panuccio, Tripepi, Cutrupi, Parlongo, Catalano, Tanaka, Ouchi, Kihara, Funahashi, Matsuzawa: "Adiponectin is markedly increased in patients with nephrotic syndrome and is related to metabolic risk factors." in: Kidney international. Supplement, Issue 84, pp. S98-102, 2003 (PubMed).

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