This product includes 1 ml of Reagent CLA and 0.1 ml of 10 mM NaCl solution. It is for measurement of 100 samples using 96-well plates. Cuvettes may also be used for measurements.
Chloride plays a variety of important physiological roles in the cells. Chloride channels are responsible for cell membrane potential and regulating cell volume. The chloride level in neurons is related to the actions of neurotransmitters glycine and GABA . The chloride level in blood is directly related to the kidney functions. Chlorides are the most common reagents and buffers used in research and the most abundant anion in nature. The MicroMolar Chloride Assay Kit (Catalog number CLA100) provides a quick and simple method for quantification of chloride in a variety of samples. The kit does not contain toxic mercury component. It is based on measurement of optical density at 360 nm (OD360) that correlates the chloride concentration in the presence of Reagent CLA. The assay linear range is 0.01 mM – 1 mM. Samples with higher chloride concentrations are diluted. The assay is compatible with a HEPES buffer.
The MicroMolar Chloride Assay Kit (catalog number CLA100) includes 3 ml of Reagent CLA and 0.1 ml of 10 mM KCl solution. It is for measurement of 100 samples using 96-well plates. Cuvettes may also be used for measurements.
ASSAY PROTOCOL
The following assay protocol is based on using a 96-well plate for the measurement. The sample volume is 150 µl and the final assay volume is 180 µl. Please adjust the sample volume and reagent volume proportionally if a 384-well plate or a cuvette is used for the measurement.
STANDARD CURVE
1. Sample preparation: Prepare 150 µl of potassium chloride solutions in the wells of a clear 96-well plate (Corning 3641) with a two-fold serial dilution from 1 mM to zero in water.
2. Detection: Mix 30 µl of Reagent CLA with 150 µl of the Chloride solutions for 20 min. Shake the plate and read the optical density at 360 nm (OD360).
3. Data Analysis: Plot the OD360 and the chloride concentration [Chloride] to generate the linear standard curve.
OD360 = a [Chloride] + b
Where the OD360 values are from experimental data, the a and b values are from the linear fitting between the OD360 values and the chloride concentrations.
UNKNOWN SAMPLES
Follow the same procedure to measure the OD360 values from the unknown samples. Calculate the chloride concentrations in the unknown samples using the OD360 values from the unknown samples and the a and b values from the standard curve.