Free Glycerol Colorimetric/Fluorometric Assay Kit

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Reaktivität
Chemical
2
Detektionsbereich
0.05-10 nM
Untere Nachweisgrenze
0.05 nM
Applikation
Biochemical Assay (BCA)
Optionen
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Proben Cell Culture Supernatant, Cell Samples, Plasma, Serum, Tissue Samples
Nachweismethode Fluorometric, Colorimetric
Spezifität Glycerol Assay Kit provides a sensitive, easy assay to measure free glycerol concentration in various samples. In the assay, glycerol is enzymatically oxidized to generate a product which reacts with the probe to generate color (lambda = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect 50 pmol-10 nmol (or 1 approx. 10000 ?M range) of glycerol in various samples.
Produktmerkmale Free Glycerol Assay Kit: Colorimetric & Fluorometric Assay to Measure free Glycerol concentration in various samples within 40 min. Fast, Easy & Sensitive.
Bestandteile Glycerol Assay Buffer
Glycerol Probe (in DMSO, anhydrous)
Glycerol Enzyme Mix (lyophilized)
Glycerol Standard (100 mM)
Substanzklasse Chemical
Hintergrund Glycerol is widely used in foods, beverages, solvents, pharmaceutical and cosmetic products, etc. There is broad interest in quantification of glycerol for research and development.
Applikationshinweise The kit can detect 50 pmol-10 nmol (or 1~10000 μM range) of glycerol in various samples.
Kommentare

Further details regarding sample type: Animal tissues & cells, Cell and tissue culture supernatants, plasma and serum, as well as many other biological fluids

Testdauer < 1 h
Protokoll 1. Standard Curve Preparation: For the colorimetric assay, add 10 µL of the glycerol standard to 990 µL of Assay Buffer to generate 1 mM glycerol standard, mix well. Add 0, 2, 4, 6, 8, 10 µL into each well individually. Adjust volume to 50 µL/well with Assay Buffer to generate 0, 2, 4, 6, 8, 10 nM/well of glycerol Standard. For the fluorometric assay, dilute the Glycerol Standard to 0.01- 0.1 mM with the Assay Buffer (Detection sensitivity is 10-100 fold higher for a fluorometric than a colorimetric assay). Follow the protocol as for the colorimetric assay.
2. Sample Preparation: Prepare test samples to a final volume of 50 µL/well with Assay Buffer in a 96-well plate. We suggest testing several dilutions of your sample to make sure the readings are within the standard curve range.
3. Reaction Mix: Mix enough reagent for the number of samples and standards to be performed: For each well, prepare a total 50 µL Reaction Mix: 46 µL Assay Buffer 2 µL Glycerol Probe 2 µL Glycerol Enzyme Mix
4. Add 50 µL of the Reaction Mix to each well containing standard and samples. Mix well. Incubate at room temperature for 30 minutes, protect from light.
5. Measure O.D. 570 nm for the colorimetric assay or Ex/Em = 535/590 nm for the fluorometric assay in a microtiter plate reader. The reaction is stable for at least two hours.
Ergebnisberechnung

Calculations: Correct background by subtracting the value derived from the 0 glycerol standard from all sample readings. Plot the standard curve (OD 570nm or Fluorescence readings vs. nmol). Apply sample readings to the standard curve. Glycerol concentration can then be calculated: C = Ga / Sv nM/µL or µM/mL or mM Where: Ga is Glycerol amount from standard curve (nmol). Sv is the sample volume (before dilution) added in sample wells (µL). Glycerol molecular weight: 92.09.

Beschränkungen Nur für Forschungszwecke einsetzbar
Lagerung -20 °C
Haltbarkeit 12 months
Produkt verwendet in: Tsao, Shiau, Chuang, Chang, Hwang: "Interleukin-4 regulates lipid metabolism by inhibiting adipogenesis and promoting lipolysis." in: Journal of lipid research, Vol. 55, Issue 3, pp. 385-97, 2014 (PubMed).

Macdonald, Wan, Frendo-Cumbo, Dyck, Wright: "IL-6 and epinephrine have divergent fiber type effects on intramuscular lipolysis." in: Journal of applied physiology (Bethesda, Md. : 1985), Vol. 115, Issue 10, pp. 1457-63, 2013 (PubMed).

Gögebakan, Andres, Biedasek, Mai, Kühnen, Krude, Isken, Rudovich, Osterhoff, Kintscher, Nauck, Pfeiffer, Spranger: "Glucose-dependent insulinotropic polypeptide reduces fat-specific expression and activity of 11β-hydroxysteroid dehydrogenase type 1 and inhibits release of free fatty acids." in: Diabetes, Vol. 61, Issue 2, pp. 292-300, 2012 (PubMed).

Yogosawa, Mizutani, Ogawa, Izumi: "Activin receptor-like kinase 7 suppresses lipolysis to accumulate fat in obesity through downregulation of peroxisome proliferator-activated receptor γ and C/EBPα." in: Diabetes, Vol. 62, Issue 1, pp. 115-23, 2012 (PubMed).

Huo, Guo, Li, Wang, Zhang, Wang, Zhou, Gao, Telang, Chesney, Chen, Ye, Chapkin, Wu: "Disruption of inducible 6-phosphofructo-2-kinase ameliorates diet-induced adiposity but exacerbates systemic insulin resistance and adipose tissue inflammatory response." in: The Journal of biological chemistry, Vol. 285, Issue 6, pp. 3713-21, 2010 (PubMed).

Pentinat, Ramon-Krauel, Cebria, Diaz, Jimenez-Chillaron: "Transgenerational inheritance of glucose intolerance in a mouse model of neonatal overnutrition." in: Endocrinology, Vol. 151, Issue 12, pp. 5617-23, 2010 (PubMed).

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