Chloride Assay Kit

Catalog No. ABIN1000258

Product Details

Target: Chloride

Application: Biochemical Assay (BCA)

Sample Type: Serum, Plasma, Urine, Saliva, Sweat, Food, Beverages, Water, Soil

Specificity: 0.7 mg/dL (0.2 mM)

Characteristics: Sensitive and accurate. Use as little as 5 µL samples. Linear detection range 0.7 mg/dL (0.2mM) to 35 mg/dL (10mM) Cl - in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 5 min. Can be readily automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvet or 96-well plate assay.
Low interference in biological samples.
No pretreatments are needed. Assays can be directly performed on raw biological samples i.e., in the presence of lipid, protein and minerals such as magnesium, iron and zinc.

Components: Reagent: 50 mL. Chloride standard: 1 mL 35 mg/dL Cl-.

Material not included: Pipeting devices and accessories (e.g. 5 µL), clear flat-bottom 96- well plates and plate reader, or spectrophotometer and cuvets.

Application Details

Application Notes: Direct Assays: Cl in serum, plasma, urine, saliva, sweat etc.
Drug Discovery/Pharmacology: effects of drugs on chloride metabolism.
Food and Beverages: chloride determination.
Environment: chloride determination in water and soil.

Protocol: Procedure using 96-well plate:
1. Dilute standards in distilled water as shown in the table. Serum, plasma, urine and milk samples should be diluted 20-fold in water. Transfer 5 µL diluted standards and samples to wells of a clear bottom 96-well plate. Store diluted standards at 4°C for future use.
2. Add 200 µL working reagent and tap lightly to mix.
3. Incubate 5 min at room temperature and read optical density at 610 nm (550-650nm nm).

Procedure using cuvette:
1. Set up test tubes labeled Standards and Samples. Transfer 10 µL diluted standards and samples to appropriately labeled tubes.
2. Add 1000 µL working reagent and vortex to mix. Incubate 5 min, transfer to cuvet and read OD at 610nm.

Reagent Preparation:

Important: bring reagents to room temperature and shake well before use.

Calculation of Results:

Subtract blank OD (water, #8) from the standard OD values and plot the OD against Cl - standard concentrations. Determine the slope using linear regression fitting.
Conversions: 1 mg/dL Cl - equals 282 µM, 0.001% or 10 ppm.

Restrictions: For Research Use only

Handling

Storage: 4 °C

References

Grünberg, Donkin, Constable: "Periparturient effects of feeding a low dietary cation-anion difference diet on acid-base, calcium, and phosphorus homeostasis and on intravenous glucose tolerance test in high-producing dairy cows." in: Journal of dairy science, Vol. 94, Issue 2, pp. 727-45, (2011) (PubMed).

Peng, Adelman, Rizzolo: "Minimal effects of VEGF and anti-VEGF drugs on the permeability or selectivity of RPE tight junctions." in: Investigative ophthalmology & visual science, Vol. 51, Issue 6, pp. 3216-25, (2010) (PubMed).

Rajapakshe, Johnson-Winters, Nordstrom, Meyers, Emesh, Astashkin, Enemark: "Characterization of chloride-depleted human sulfite oxidase by electron paramagnetic resonance spectroscopy: experimental evidence for the role of anions in product release." in: Biochemistry, Vol. 49, Issue 25, pp. 5154-9, (2010) (PubMed).

Everaert, Willemsen, Kamers, Decuypere, Bruggeman: "Regulatory capacities of a broiler and layer strain exposed to high CO2 levels during the second half of incubation." in: Comparative biochemistry and physiology. Part A, Molecular & integrative physiology, Vol. 158, Issue 2, pp. 215-20, (2010) (PubMed).

Ahmad, Mushtaq, Khan, Babar, Yousaf, Hasan, Kamran: "Influence of varying dietary electrolyte balance on broiler performance under tropical summer conditions." in: Journal of animal physiology and animal nutrition, Vol. 93, Issue 5, pp. 613-21, (2009) (PubMed).

Borenshtein, Schlieper, Rickman, Chapman, Schweinfest, Fox, Schauer: "Decreased expression of colonic Slc26a3 and carbonic anhydrase iv as a cause of fatal infectious diarrhea in mice." in: Infection and immunity, Vol. 77, Issue 9, pp. 3639-50, (2009) (PubMed).

Jensen, Mouridsen, Reibel, Brünner, Nauntofte: "Adjuvant chemotherapy in breast cancer patients induces temporary salivary gland hypofunction." in: Oral oncology, Vol. 44, Issue 2, pp. 162-73, (2008) (PubMed).

Target Details

Target: Chloride

Target Type: Anorganic

Background: Quantitative determination of chloride ion Cl- by colorimetric (610nm) method.
Procedure: 5 min.

Chloride is the major extracellular anion in human body fluids. Chloride plays a key role in maintaining proper water distribution, osmotic pressure and electrolyte balance. Low chloride concentrations may be found with prolonged vomiting, extensive burns, metabolic acidosis, Addisonia crisis and renal diseases. Elevated chloride concentrations are associated with dehydration, congestive heart failure, hyperventilation and urinary obstructions. Determination of chloride in sweat is useful in diagnosing cystic fibrosis. Simple, direct and automation-ready procedures for measuring chloride concentration in biological samples are becoming popular in Research and Drug Discovery. This chloride assay kit is designed to measure chloride directly in biological samples without any pretreatment. The improved Fried method utilizes mercuric 2,4,6-tripyridyl-s-triazine, which forms a colored complex specifically with chloride. The intensity of the color, measured at 610nm, is directly proportional to the chloride concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples.