Sulfate Assay Kit, BioAssay(TM)

Produktübersicht

• Artikelname: Sulfate Assay Kit, BioAssay(TM)
• Artikelnummer: USB-S8016-96
• Hersteller Artikelnummer: S8016-96
• Alternativnummer: USB-S8016-96-96
• Hersteller: US Biological
• Kategorie: Kits/Assays

Produktbeschreibung

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 osteochon- drodysplasia 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. 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. Key Features: Sensitive and accurate. Detection range 0.02mM (0.19 mg/dL) to 2mM (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. Applications: 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: 25ml Reagent B: 2.4 g Powder TCA Reagent: 25ml Sulfate Standard: 1ml 60mM 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. Please refer to Material Safety Data Sheet for detailed information. Procedures: Sample Treatment: Urine samples should be diluted 10-fold in deionized water prior to assay. Fresh serum or plasma (non-hemolyzed) samples can be either assayed immediately, or frozen for future tests. Samples should be deproteinated as follows: mix 200ul sample and 100ul TCA Reagent in a 1.5-mL Eppendorf tube. Spin down protein precipitates 5 min at 14,000 rpm on a table centrifuge. Transfer 200ul supernatant for assay. Procedure using 96-well plate: 1. Standards. Prepare a 2.0mM Premix by mixing 20ul 60mM Sulfate Standard with 580ul dH2O. Dilute Premix as follows. No Premix+H2O Sulfate (mM) 1 200ul+10ul 2.0 2 100ul+100ul 1.0 3 50ul+150ul 0.5 4 0ul+200ul 0 Transfer 200ul Standards into separate wells of a clear flat-bottom 96-well plate. Note: when deproteination is required (i.e. serum or plasma), treat the standards by adding 100ul TCA Reagent to 200ul of each standard, mix and transfer 200ul of the resulting standard into separate wells. Transfer 200ul Samples (see above for Sample Treatment) into separate wells of the plate. 2. Working Reagent. The Working Reagent (WR) must be prepared fresh and used within 1 hour after reconstitution. Prepare enough WR for all samples and standards (100ul WR per assay well) by mixing 95 mg Reagent B perml Reagent A, i.e. 10 assay wells, mix 95 mg Reagent B with 1ml Reagent A. Vortex for at least 1 min to ensure complete dissolution of the powder and incubate the reconstituted Working Reagent for 10 min before use. Use a multi-channel pipettor, add 100ul Working Reagent to each assay well. Tap plate to mix well. If TCA precipitation was required, mixing the samples and standards with the WR can be improved by pipetting up and down once. 3. Incubate 5 min at room temperature and read optical density at 540- 610nm (600nm). Calculation: Sulfate concentration (mM) in the sample can be calculated as follows, [Sulfate]=ODSAMPLE-ODH2O * n (mM) Slope ODSAMPLE and ODH2O are the OD values of the sample well and the water well (Standard 4), respectively. n=10 for urine. Conversions: 1mM sulfate equals 9.61 mg/dL or 96.1 ppm. G