Ascorbic Acid Colorimetric/Fluorometric Assay Kit

Details zu Produkt Nr. ABIN411633, Anbieter: Anmelden zum Anzeigen
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Reaktivität
Chemical
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Detektionsbereich
0.01-10 nM
Untere Nachweisgrenze
0.01 nM
Applikation
Biochemical Assay (BCA)
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Verwendungszweck Ascorbic Acid Assay Kit provides a rapid, simple, and sensitive means of detecting ascorbic acid in various biological samples. In this assay, our proprietary catalyst oxidizes ascorbic acid to produce a product that interacts with the ascorbic acid probe, generating color and fluorescence. Ascorbic acid can be easily determined by either colorimetric (spectrophotometry at λ = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods. The assay can detect 0.01-10 nmol of ascorbic acid per assay in various samples.
Proben Cell Culture Supernatant, Plasma, Serum, Tissue Samples
Nachweismethode Fluorometric, Colorimetric
Spezifität Ascorbic Acid Assay Kit provides a rapid, simple, and sensitive means of detecting ascorbic acid in various biological samples. In this assay, our proprietary catalyst oxidizes ascorbic acid to produce a product that interacts with the ascorbic acid probe, generating color and fluorescence. Ascorbic acid can be easily determined by either colorimetric (spectrophotometry at lambda = 570 nm) or fluorometric (Ex/Em = 535/587 nm) methods. The assay can detect 0.01-10 nmol of ascorbic acid per assay in various samples.
Produktmerkmale Ascorbic Acid Assay Kit: Colorimetric & Fluorometric Assay for detecting Ascorbic Acid in various Biological Samples such as Serum etc. within 5 min. Convenient, Accurate & Reliable.
Bestandteile Ascorbic Acid Assay Buffer
Ascorbic Acid Probe (in DMSO)
Catalyst
Ascorbic Acid Enzyme Mix (lyophilized)
Ascorbic Acid Standard (20 μmole)
Substanzklasse Chemical
Hintergrund Ascorbic Acid (Vitamin C) plays an important role in many biological processes. It is a potent anti-oxidant, anti-inflammatory, anti-viral agent, and an immune stimulant and is present is a wide variety of foods and biological specimens. It is important to be able to monitor ascorbic acid content in these different samples.
Applikationshinweise The assay can detect 0.01-10 nmol of ascorbic acid per assay in various samples.
Kommentare

Further details regarding sample type: Cell and tissue culture supernatants, urine, plasma and serum, as well as many other biological fluids, food samples, growth medium, etc.

Testdauer 5 min
Protokoll 1. Standard Curve Preparations: For the colorimetric assay, dilute the standard to 1 mM by adding 10 µL of the 100 mM Ascorbic Acid Standard to 990 µL of distilled water, mix well. Add 0, 2, 4, 6, 8, 10 µL into each well individually. Adjust volume to 120 µL/well with Ascorbic Acid Assay Buffer to generate 0, 2, 4, 6, 8, 10 nM/well of Ascorbic Acid Standard. For the fluorometric assay, dilute the Ascorbic Acid Standard to 0.01- 0.1 mM with the Ascorbic Acid Assay Buffer (Note: Detection sensitivity is 10 to 100 fold higher for a fluorometric than a colorimetric assay). Follow the procedure for the colorimetric assay. Note: Diluted ascorbic acid standard is unstable, use fresh dilution each time.
2. Sample Preparation: Prepare test samples to a final volume of 120 µL/well with Ascorbic Acid Assay Buffer in a 96-well plate. We suggest testing several doses of your sample to make sure the readings are within the standard curve range. NOTE: 1) Due to high protein contents and other compounds present in serum we recommend using FRASC Ascorbic Acid Kit (ABIN411634) for serum samples. 2) Ascorbate is easily oxidized during sample preparation and great care must be exercised to achieve quantitative recovery.
3. Catalyst: Add 100 µL of catalyst to 900 µL of distilled water and vortex well.
4. Add 30 µL of catalyst to each standard and sample well.
5. Ascorbic Acid Reaction Mix: Mix enough reagents for the number of samples and standards to be performed: For each well, prepare a total 50 µL Reaction Mix containing: 46 µL Ascorbic Acid Assay Buffer 2 µL Ascorbic Acid Probe 2 µL Ascorbic Acid Enzyme Mix
6. Mix well. Add 50 µL of the Reaction Mix to each well containing the Ascorbic Acid Standard and test samples. Mix well.
7. Protect from light, Color is developed within 3 minutes and stable for an hour.
8. Measure O.D. 570nm for colorimetric assay or Ex/Em = 535/590 nm for fluorometric assay in a micro-plate reader.
9. Correct background by subtracting the value derived from the 0 ascorbic acid standard from all sample readings (Note: The background reading can be significant and must be subtracted from sample readings). Apply sample readings to the generated standard curve. Ascorbic Acid concentration can then be calculated: C = As / Sv nM/µL or µM/mL or mM Where: As is ascorbic acid amount from standard curve (nmol). Sv is the sample volume added in sample wells (µL). Ascorbic Acid molecular weight: 176.12.
Beschränkungen Nur für Forschungszwecke einsetzbar
Lagerung -20 °C
Haltbarkeit 12 months
Produkt verwendet in: Cui, Pan, Zhang, Jones, Zhang: "Progressive pseudogenization: vitamin C synthesis and its loss in bats." in: Molecular biology and evolution, Vol. 28, Issue 2, pp. 1025-31, 2011 (PubMed).

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