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#DSFT-200 QuantiChrom™ Sulfate Assay Kit
Product name : QuantiChrom™ Sulfate Assay Kit
Catalog number : DSFT-200
Supplier name : Bioassays
Data sheet: Ask more or other datasheet now !
About this Product :QuantiChrom™ Sulfate Assay Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of QuantiChrom™ Sulfate Assay Kit.
More Details about
For quantitative determination of sulfate ion and evaluation of drug effects on sulfate metabolism.
Samples: serum, urine, food and environment.
Procedure: 5 min.
Size: 200 tests.
Detection limit: 20 μM.
INORGANIC SULFATE is one of the most abundant anions in
mammalian plasma. Sulfate plays important physiological roles in
activating and detoxifying xenobiotics, steroids, neurotransmitters, and
bile acids. Sulfate is needed for the biosynthesis of glycosaminoglycans,
cerebroside sulfate, and heparin sulfate. Undersulfation of cartilage
proteoglycans has been associated with human inherited osteochondrodysplasia
disorders. In mammals, sulfate homeostasis is regulated by
the kidney. The majority of filtered sulfate is absorbed in the proximal
tubules, and only 5–20% of the filtered load is excreted into the urine.
Simple, direct and automation-ready procedures for quantitative
determination of inorganic sulfate find wide applications in research and
drug discovery. BioAssay Systems' sulfate assay kit is designed to
measure sulfate concentration in biological fluids such as serum and
urine. The improved method utilizes the quantitative formation of
insoluble barium sulfate in polyethylene glycol. The turbidity measured
between 540 and 610nm is proportional to sulfate level in the sample.
Sensitive and accurate. Detection range 0.02 mM (0.19 mg/dL) to 2 mM
(19.2 mg/dL) sulfate in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a
single working reagent and incubation for 5 min.
Direct Assays: inorganic sulfate in serum and urine.
Pharmacology: effects of drugs on sulfate metabolism.
KIT CONTENTS (200 tests in 96-well plates)
Reagent A: 25 mL Reagent B: 2.4 g Powder
TCA Reagent: 25 mL Sulfate Standard: 1 mL 60 mM
Storage Conditions. The kit is shipped and stored at room temperature.
Shelf life of three months after receipt.
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents.
MATERIALS REQUIRED, BUT NOT PROVIDED
Pipetting devices and accessories, clear flat-bottom 96-well plates and
plate reader, or cuvets and spectrophotometer.
Sample Type: Sulfate content of polysaccharides from ray skin
Species: Raja radula
References: Ben Mansour, M et al (2009). Polysaccharides from the skin of the ray Raja radula. Partial characterization and anticoagulant activity. Thromb Res 123(4):671-8.
Pubmed ID: 18617224
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=18617224
INTRODUCTION: The polysaccharide fraction from the skin of the ray Raja radula was extracted, characterized and assayed for anticoagulant activity.
MATERIALS AND METHODS: A whole polysaccharidic fraction was extracted from the skin of the ray Raja radula by papain digestion followed by cetylpyridinium chloride and ethanol precipitation and was subjected to gel chromatography and anion exchange chromatography, acetate cellulose electrophoresis and characterized by physicochemical procedures. APTT and anti Xa assays were performed to assess the anticoagulant activity of the polysaccharidic fractions in comparison with unfractionated heparin.
RESULTS: Gel and anion-exchange chromatography revealed two negatively charged polysaccharidic populations different in both molecular weight and charge. Infrared spectra suggested the occurrence of uronic acids and acetylated hexosamines. The second polysaccharide was highly sulfated, with a sulfate content of approximately 29%. These data suggested that dermatan sulfate (DS) is the sulfate rich polysaccharide whereas hyaluronic acid (HA) is the polysaccharide devoid of sulfate groups. Molecular mass characterization indicated that their average molecular masses were 22 kDa and 85 kDa, respectively. The sulfated polysaccharide, i.e. presumably DS, accounted alone for the observed concentration-dependent anticoagulant activity which was, as measured by APTT, 2 to 3-fold lower than that of heparin. In addition, it had a significant anti-Xa activity.
CONCLUSION: A major-sulfated polysaccharide, likely a dermatan sulfate, was extracted from the ray Raja radula skin. The results indicated that it exhibited a high anticoagulant activity and suggested that it was mediated by both heparin cofactor II and antithrombin. [PubMed - indexed for MEDLINE]
Sample Type: tissue
Species: Raja radula
References: Ben Mansour, M et al (2010). Highly sulfated dermatan sulfate from the skin of the ray Raja montagui: anticoagulant activity and mechanism of action. Comp Biochem Physiol B Biochem Mol Biol 156(3):206-15.
Pubmed ID: 20363356
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=20363356
Abstract: The dermatan sulfate (DS) isolated from the ray skin Raja montagui was identified and characterized. Its average molecular weight (Mw) and sulfate content were 39 kDa and 25% w/w, respectively. This DS prolonged thrombin time and activated partial thromboplastin time and inhibited the thrombin generation in a concentration-dependent manner whereas it had no effect on the anti-Xa assay and on platelet function. Data from the anti-IIa assay allowed the assessment of the specific anticoagulant activity which was 40 units/mg. The kinetics of the thrombin inhibition by heparin cofactor II (HCII) has been studied as a function of DS concentration according to a kinetic model in which the polysaccharide binds quickly to the inhibitor and forms a complex more reactive than the free inhibitor towards thrombin. This DS accelerated thrombin inhibition exclusively by HCII. The dissociation constant of the DS-HCII complex, K(DSHCII), and the rate constant of the thrombin inhibition by this complex, k, were (2.93+/-0.25)x10(-6)M and (2.2+/-0.35)x10(9)M(-1)min(-1), respectively. Our findings indicated that the major polysaccharide in the skin of the ray Raja montagui was a DS endowed with a high anticoagulant effect mediated by HCII and which may constitute an anticoagulant drug of interest in anticoagulant therapy.
(c) 2010 Elsevier Inc. All rights reserved.[PubMed - indexed for MEDLINE]
References: Ben Mansour, M et al (2009). Characterization of a novel dermatan sulfate with high antithrombin activity from ray skin (Raja radula). Thromb Res 123(6):887-94.
Pubmed ID: 19019412
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=19019412
A novel dermatan sulfate (DS) from the skin of the ray Raja radula with high anticoagulant activity was identified and its monosaccharide composition and anticoagulant mode of action and potency were determined.
MATERIALS AND METHODS:
The DS isolated from the ray skin was identified by chondroitinase treatment and characterized by FT-IR and (1)H NMR spectroscopy. Its anticoagulant activity was checked by activated partial thromboplastin time (aPTT), thrombin time (TT), thrombin generation (TG), heparin cofactor II (HCII) and antithrombin (AT)-mediated inhibition of thrombin. The effects on platelet activation and aggregation were investigated using flow cytometry and aggregometry, respectively.
Chemical backbone structures of DS from Raja radula were close to that of DS from porcine intestinal mucosa. However, (1)H NMR indicated that iduronic acid was the major hexuronic acid moiety in the ray skin DS and also suggested that the amount of 2-O-sulfonated iduronic acid was higher in comparison with mammalian DS along with the occurrence of 4-O-sulfonated N-acetylgalactosamine residues. The anticoagulant effect of the ray skin DS was mainly due to the potentiation of thrombin inhibition by HCII but also, although to a lesser extent, by AT and was higher than that of the DS standard. Moreover, it had no effect on platelet activation and aggregation induced by various agonists.
Altogether, these results indicated that DS from raja radula skin is an anticoagulant drug of interest potentially useful in anticoagulant therapy.
[PubMed - indexed for MEDLINE]
Sample Type: culture medium
References: Majdoub, H et al (2009). Anticoagulant activity of a sulfated polysaccharide from the green alga Arthrospira platensis. Biochim Biophys Acta 1790(10):1377-81.
Pubmed ID: 19632306
Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed?term=19632306
BACKGROUND: The polysaccharide of culture medium from Arthrospira platensis was extracted by ultrafiltration, partially characterized and assayed for anticoagulant activity.
METHODS: The crude polysaccharidic fraction was fractionated by anion exchange chromatography on DEAE-cellulose, subjected to acetate cellulose electrophoresis and characterized by physicochemical procedures. The anticoagulant effect of the ultrafiltrated polysaccharide was checked by several coagulation tests.
RESULTS: Anion exchange chromatography revealed in the whole ultrafiltrated polysaccharidic fraction the occurrence of a sulfated spirulan-like component designated PUF2. The average molecular weight of PUF2 was determined by size exclusion chromatography combined with multi-angle light scattering (SEC-MALS) and viscosimetry and was 199 kDa and the sulfate content was 20% weight/dry weight. The physicochemical characterization indicated the occurrence of rhamnose (49.7%), galacturonic and glucuronic acid (32% of total sugar). The anticoagulant effect of this sulfated polysaccharide was mainly due to the potentiation of thrombin inhibition by heparin cofactor II and was 4-times higher than that of the porcine dermatan sulfate whereas it had no effect on anti-Xa activity.
CONCLUSIONS: An ultrafiltrated sulfated polysaccharide, likely a calcium spirulan was obtained from the culture medium of A. platensis and showed an anticoagulant activity mediated by heparin cofactor II.
Old culture medium of A. platensis may represent an important source for the spirulan-like PUF2 which was endowed with potentially useful anticoagulant properties whereas its obtention by ultrafiltration may represent an extraction procedure of interest.
[PubMed - indexed for MEDLINE]
Sample Type: Sulfate content of exopolysaccharides from ascomycete fungus
Species: Cordyceps sinensis
References: Wang, ZM et al (2011). Structural characterisation and immunomodulatory property of an acidic polysaccharide from mycelial culture of Cordyceps sinensis fungus Cs-HK1. Food Chemistry 125(2):637-43.
Pubmed ID: n/a
Pubmed link: n/a
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