Sialic Acid Assay Kit

Produktnummer ABIN1000330

Produktdetails

Target: Sialic Acid (SA)

Applikation: Biochemical Assay (BCA)

Produktmerkmale: Sensitive and accurate. Use as little as 10 µL samples. Linear detection range in 96-well plate: 0.02 to 1 mM sialic acid for colorimetric assays and 2 to 100 µM for fluorimetric assays.
Simple and convenient. Can detect free sialic acid by addition of a single working reagent and incubation for 60 min at room temperature or total sialic acid by pre-treating samples with a 60 min hydrolysis step.

Bestandteile: Assay Buffer: 10 mL. Hydrolysis Reagent: 10 mL. Enzyme: 120 µL. Neutralization Reagent: 5 mL. Dye Reagent: 120 µL. Standard: 500 µL 10 mM Sialic Acid.

Benötigtes Material: Pipeting devices, centrifuge tubes, Clear flat-bottom 96-well plates, black 96-well or 384-well plates (e.g. Corning Costar) and plate reader.

Anwendungsinformationen

Applikationshinweise: Direct Assays: sialic acid in biological samples.

Protokoll: Note: For measurement of free sialic acid, this procedure should be skipped.
1. Combine 20 µL of sample with 80 µL Hydrolysis Reagent in a microcentrifuge tube (screw cap tube is preferable) and incubate at 80°C for 60 min.
2. Allow sample to cool to room temperature and briefly centrifuge at 14000 rpm to spin down the hydrolysis mixture.
3. Add 20 µL Neutralization Reagent to each hydrolysis reaction, briefly vortex to mix and briefly centrifuge at 14000 rpm to spin down the reaction. The samples are now ready for the sialic acid assay.

Ergebnisberechnung:

Subtract blank OD (water, #4) from the standard OD values and plot the OD against standard concentrations.
Conversions: 1 mM NANA equals 30.9 mg/dL or 309 ppm. FLUORIMETRIC

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -20 °C

Referenzen

Chrostek, Supronowicz, Panasiuk, Cylwik, Gruszewska, Szmitkowski: "Serum Sialic Acids Levels According to the Severity of Liver Cirrhosis." in: Journal of clinical laboratory analysis, (2014) (PubMed).

Park, Kim, Ahn, Lee, Kim, Kim, Woo: "Structural and biochemical characterization of the broad substrate specificity of Bacteroides thetaiotaomicron commensal sialidase." in: Biochimica et biophysica acta, Vol. 1834, Issue 8, pp. 1510-9, (2013) (PubMed).

Bonten, Yogalingam, Hu, Gomero, van de Vlekkert, dAzzo: "Chaperone-mediated gene therapy with recombinant AAV-PPCA in a new mouse model of type I sialidosis." in: Biochimica et biophysica acta, Vol. 1832, Issue 10, pp. 1784-92, (2013) (PubMed).

Cylwik, Chludzinska, Chrostek, Gruszewska, Gindzienska-Sieskiewicz, Szmitkowski, Sierakowski: "Relationship between CDT and disease activity in rheumatoid arthritis." in: Zeitschrift für Rheumatologie, Vol. 71, Issue 3, pp. 220-3, (2012) (PubMed).

Cylwik, Chrostek, Brodowska-Adamusiak, Gruszewska, Daniluk, Szmitkowski: "The changes of sialic acid concentration and content in apolipoprotein B-containing lipoproteins in the sera of alcoholics." in: Alcohol and alcoholism (Oxford, Oxfordshire), Vol. 45, Issue 5, pp. 422-6, (2010) (PubMed).

Antigendetails

Target: Sialic Acid (SA)

Andere Bezeichnung: Sialic Acid (SA Produkte)

Hintergrund: Quantitative determination of free and total sialic acid (NANA) by colorimetric (570nm) or fluorimetric (530nm/590nm) methods.
Procedure: 2 hrs.

Sialic Acid is a general name for nine carbon acidic sugars with N- or O-substituted derivatives. The most common member of these sugars is N-acetylneuraminic acid (NANA). Sialic acid is widely distributed throughout mammalian tissues and fluids including serum. Sialylated oligosaccharides have been shown to exhibit antiviral properties and are also known to influence blood coagulation and cholesterol levels. The Sialic acid level in body fluids is also an important marker for diagnosing cancer. Simple, direct and automation-ready procedures for measuring sialic acid concentrations find wide applications in research and drug discovery. This sialic acid assay uses a single Working Reagent that combines NANA aldolase, pyruvate oxidase and hydrogen peroxide determination in one step. The color intensity of the reaction product at 570nm or fluorescence intensity at gamma em/ex = 585/530nm is directly proportional to sialic acid concentration in the sample.