Sulfate Assay Kit

Catalog No. ABIN1000276

Product Details

Target: Sulfate

Application: Biochemical Assay (BCA)

Sample Type: Serum, Urine

Specificity: 10 μM

Characteristics: 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.

Components: Reagent A: 25 mL. Reagent B: 2.4 g. Powder TCA Reagent: 25 mL. Sulfate Standard: 1 mL 60 mM.

Material not included: Pipetting devices and accessories, clear flat-bottom 96-well plates and plate reader, or cuvets and spectrophotometer.

Application Details

Application Notes: Direct Assays: inorganic sulfate in serum and urine.
Pharmacology: effects of drugs on sulfate metabolism.

Comment:

This procedure can be scaled up to assays in a cuvette. The following compounds have been tested and do not interfere: 400 mM sodium chloride, 500 mM urea, 5 mM sodium phosphate, 4 mM sodium citrate, 1.5 mM sodium EDTA. Ester sulfate can be determined following a digestion step (Lundquis, P. et al. (1980). Clin. Chem. 26:1178-81). This will allow quantification of total inorganic and ester sulfate in a sample.

Protocol: Procedure using 96-well plate:
1. Standards. Prepare a 2.0 mM Premix by mixing 20 µL 60 mM Sulfate Standard with 580 µL dH2O. Dilute Premix. Transfer 200 µL 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 100 µL TCA Reagent to 200 µL of each standard, mix and transfer 200 µL of the resulting standard into separate wells. Transfer 200 µL 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 (100 µL WR per assay well) by mixing 95 mg Reagent B per mL Reagent A, i.e. 10 assay wells, mix 95 mg Reagent B with 1 mL 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 100 µL 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).

Sample Preparation:

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 : mix 200 µL sample and 100 µL TCA Reagent in a1.5-mL Eppendorf tube. Spin down protein precipitates 5 min at 14,000 rpm on a table centrifuge. Transfer 200 µL supernatant for assay.

Calculation of Results:

Conversions: 1 mM sulfate equals 9.61 mg/dL or 96.1 ppm.

Restrictions: For Research Use only

Handling

Storage: -20 °C

References

de Gouw, Hermans, Bootsma, Zomer, Heuvelman, Diavatopoulos, Mooi: "Differentially expressed genes in Bordetella pertussis strains belonging to a lineage which recently spread globally." in: PLoS ONE, Vol. 9, Issue 1, pp. e84523, (2014) (PubMed).

Neff, Beck, Koeman, Boguslawski, Kefene, Borgman, Ruhe: "Partial deletion of the sulfate transporter SLC13A1 is associated with an osteochondrodysplasia in the Miniature Poodle breed." in: PLoS ONE, Vol. 7, Issue 12, pp. e51917, (2013) (PubMed).

Bose, Rogers, Adams, Joye, Girguis: "Geomicrobiological linkages between short-chain alkane consumption and sulfate reduction rates in seep sediments." in: Frontiers in microbiology, Vol. 4, pp. 386, (2013) (PubMed).

Reiser, Smith, Xue, Kurtz, Liu, Legrand, He, Yu, Wong, Hinchcliffe, Tanen, Lazar, Zieba, Ichetovkin, Chen, ONeill, Tanaka, Marton, Liao, Morris, Hailman, Tokiwa, Plump: "High-throughput simultaneous analysis of RNA, protein, and lipid biomarkers in heterogeneous tissue samples." in: Clinical chemistry, Vol. 57, Issue 11, pp. 1545-55, (2011) (PubMed).

Ben Mansour, Dhahri, Hassine, Ajzenberg, Venisse, Ollivier, Chaubet, Jandrot-Perrus, Maaroufi: "Highly sulfated dermatan sulfate from the skin of the ray Raja montagui: anticoagulant activity and mechanism of action." in: Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology, Vol. 156, Issue 3, pp. 206-15, (2010) (PubMed).

Sun, Yang, Tiegs, Arend, McGraw, Penn, Petrovic: "Deletion of the pH sensor GPR4 decreases renal acid excretion." in: Journal of the American Society of Nephrology : JASN, Vol. 21, Issue 10, pp. 1745-55, (2010) (PubMed).

Ben Mansour, Majdoub, Bataille, Roudesli, Hassine, Ajzenberg, Chaubet, Maaroufi: "Polysaccharides from the skin of the ray Raja radula. Partial characterization and anticoagulant activity." in: Thrombosis research, Vol. 123, Issue 4, pp. 671-8, (2009) (PubMed).

Ben Mansour, Dhahri, Bertholon, Ollivier, Bataille, Ajzenberg, Hassine, Jandrot-Perrus, Chaubet, Maaroufi: "Characterization of a novel dermatan sulfate with high antithrombin activity from ray skin (Raja radula)." in: Thrombosis research, Vol. 123, Issue 6, pp. 887-94, (2009) (PubMed).

Majdoub, Ben Mansour, Chaubet, Roudesli, Maaroufi: "Anticoagulant activity of a sulfated polysaccharide from the green alga Arthrospira platensis." in: Biochimica et biophysica acta, Vol. 1790, Issue 10, pp. 1377-81, (2009) (PubMed).

Target Details

Target: Sulfate

Background: Quantitative determination of sulfate ion by colorimetric (600nm) method.
Procedure: 5 min.

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. This 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.