Chloride Assay Kit

Antigen: Chloride

Applikation: Biochemisches Assay (BCA)

Produktnummer: ABIN411646

Produktbeschreibung

Beschreibung: Chloride is the anionic form of chlorine. It is the most common of the anions found in living organisms. Chloride ions play a variety of important physiological roles. Chloride channels are found in a variety of cells and are responsible for setting resting cell membrane potential and regulating cell volume. In the nervous system, the action of glycine and GABA are related to chloride levels in specific neurons. Chloride is also instrumental in maintaining the acid-base balance in blood. The kidneys are instrumental in closely regulating serum chloride levels. There are a number of pathologies associated with defective chloride transport, the most well-known being Cystic Fibrosis, caused by a mutation in CFTR a membrane chloride transporter. BioVision’s Chloride Assay Kit provides a quick, simple method for quantification of Chloride in a variety of biological samples. Blood and urine can be used directly after dilution with water. The assay is based upon the competition of Hg ++ and Fe ++ for TPTZ. The preferred Hg-TPTZ adduct exhibits no color. In the presence of Chloride, Hg ++ forms HgCL 2 freeing up TPTZ which then binds the available Fe ++ giving a very intense absorbance with a λ max ~ 620nm. The assay is linear in the range 20 to 120 nmol Chloride/well with detection sensitivity ~0.4 mM chloride.

Anwendungen

Protokoll: 1. Standard Curve Preparations: Add 0, 2, 4, 6, 8, 10 μl of the 10 mM Chloride standard to a series of wells. Adjust volume to 50 μl/well with water to generate 0, 20, 40, 60, 80 and 100 nmol per well of the Chloride Standard. 2. Sample Preparation: Sample Chloride concentrations can vary over a rather wide range. Urine and serum samples should be diluted 10-100X. Take 10-50 µl samples and adjust the well volume to 50 μl with dH 2 O. For unknown samples, it may be necessary to test several different amounts of sample to ensure the readings are within the standard curve. 3. Development: Add 200 µl of the Chloride Reagent to each well containing Chloride Standard or test samples. 4. Incubate at room temperature for 15 minutes. 5. Reading: Read OD at 620 nm. The signals are stable for many hours. 6. Calculation: Subtract the 0 Chloride OD reading from all standard and sample readings. This corrects for absorbance due to buffer or plate. Plot the Chloride standard curve for the 0 corrected Chloride standards (nmol/well vs. standard readings). Apply corrected sample readings (E) to the standard curve to get the amount of Chloride in the sample wells.

Beschränkungen: Nur für Forschungszwecke einsetzbar