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Sialic Acid (NANA) Colorimetric/Fluorometric Assay Kit Kit

BCA Reactivity: Chemical Fluorometric, Colorimetric Cell Culture Supernatant, Plasma, Serum, Tissue Samples
Catalog No. ABIN411705
$455.00
Plus shipping costs $45.00
100 tests
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Delivery in 2 to 3 Business Days
  • Target
    Sialic Acid (SA)
    Reactivity
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Chemical
    Detection Method
    Fluorometric, Colorimetric
    Detection Range
    0.1-10 nM
    Minimum Detection Limit
    0.1 nM
    Application
    Biochemical Assay (BCA)
    Sample Type
    Cell Culture Supernatant, Plasma, Serum, Tissue Samples
    Specificity
    Sialic Acid Assay Kit provides a simple and convenient means of measuring free Sialic Acid in a variety of biological samples. The kit utilizes an enzyme coupled reaction in which free sialic acid is oxidized resulting in development of the Oxi-Red probe to give fluorescence (Ex/Em=535/587 nm) and absorbance (O.D.=570 nm). The kit measures sialic acid in the linear range of 0.1 to 10 nmol with a detection sensitivity approx. 1 ?M concentration.
    Characteristics
    Sialic Acid (NANA) Assay Kit: Colorimetric & Fluorometric Assay to Measure free Sialic Acid in a variety of Biological Samples within 40 min. Simple & Convenient.
    Components
    Sialic Acid Assay Buffer
    Sialic Acid Probe in DMSO
    Sialic Acid Converting Enzyme
    Sialic Acid Development Mix
    Sialic Acid Standard (10 μmol)
  • Application Notes
    The kit measures sialic acid in the linear range of 0.1 to 10 nmol with a detection sensitivity ~1 μM concentration.
    Comment

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

    Assay Time
    < 1 h
    Protocol
    1. Standard Curve: For the Colorimetric Assay: Dilute 10 µL of the 100 mM Sialic Acid Standard with 990 µL dH2O to generate 1 mM standard Sialic Acid. Add 0, 2, 4, 6, 8, 10 µL of the diluted Sialic Acid Standard into a 96-well plate to generate 0, 2, 4, 6, 8, 10 nM/well standard. Bring the volume to 50 µL with Assay Buffer. For the Fluorometric Assay: Dilute the standard to 0.1 mM (0.1 nM/µL), then follow the same protocol as colorimetric assay. Will give 0, 0.2, 0.4, 0.6, 0.8, 1 nM/well Standard.
    2. Sample Preparation: Samples can be tested for free sialic acid or hydrolyzed to measure bound sialic acid as well. There are a variety of hydrolysis protocols, and users should be cautious in selecting which protocol to use. Once the sample has been prepared, add samples to a 96-well plate and bring the volume to 50 µL/well with Assay Buffer. We suggest testing several doses of your sample to make sure the readings are within the standard curve range.
    3. Reaction Mix: Mix enough reagents for the number of assays to be performed. For each well, prepare a total 50 µL Reaction Mix containing: Sialic Acid Measurement Background Control* Assay Buffer 40 µL 44 µL Sialic Acid Converting Enzyme 2 µL ---------- Sialic Acid Development Mix 2 µL 2 µL Sialic Acid Probe** 2 µL 2 µL *Pyruvate will generate background. If significant amount of pyruvate is suspected in your sample, you may do a background control. Do pyruvate background control without Sialic Acid Converting Enzyme, which will detect only pyruvate, but not Sialic Acid. The pyruvate background should be subtracted from Sialic Acid. ** For the fluorescent assay, dilute the probe 10X to reduce background reading. Add 50 µL of the Reaction Mix to each well containing the Sialic acid standard and test samples. Mix well. Incubate the reaction for 30 min at room temperature, protect from light.
    4. Measure O.D. at 570 nm, or fluorescence at Ex/Em 535/587 nm in a microplate reader.
    Calculation of Results

    Calculation: Correct background by subtracting the value derived from the 0 Sialic Acid control from all sample and standard readings (The background reading can be significant and must be subtracted from sample readings). Plot Sialic Acid standard curve. Apply sample readings to the standard curve. Sialic Acid concentrations of the test samples can then be calculated: C = S a /S v (nM/µL, or mM) where S a is the Sialic acid content of unknown samples (in nmol) from standard curve, S v is sample volume (in µL) added into the assay wells. Sialic acid Molecular Weight is 309.3 g/mol.

    Restrictions
    For Research Use only
  • Storage
    -20 °C
    Expiry Date
    12 months
  • Target
    Sialic Acid (SA)
    Alternative Name
    Sialic Acid
    Target Type
    Chemical
    Background
    Sialic acid is a generic term for the N- or O-substituted derivatives of neuraminic acid, a monosaccharide with a nine-carbon backbone. It is also the name for the most common member of this group, N-acetylneuraminic acid. Sialic acids are found widely distributed in animal tissues and to a lesser extent in other species ranging from plants and fungi to yeasts and bacteria, mostly in glycoproteins and gangliosides. It has been shown recently that sialic acid level may be associated with developmental and pathological stages.
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