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Chloride Colorimetric Assay Kit

WARNING: This product can expose you to chemicals including Hg(NO3)2, which is [are] known to the State of California to cause birth defects or other reproductive harm.  For more information go to
Catalog #: K530

Product Details

Cat # +Size K530-100
Size 100 assays
Detection Method Absorbance (620 nm)
Species Reactivity Mammalian
Applications The assay is linear in the range 20 to 120 nmol Chloride/well with detection sensitivity ~0.4 mM chloride.
Features & Benefits • Simple procedure; takes less than ~20 minutes
• Fast and convenient
•Mn²ᶧ, Cu²ᶧ, Zn²ᶧ do not interfere with the assay.
Kit Components • Chloride Reagent
• Chloride Standard (10 µmol)
Storage Conditions RT
Shipping Conditions RT
USAGE For Research Use Only! Not For Use in Humans.


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 Hg²ᶧ and Fe²ᶧ for TPTZ. The preferred Hg-TPTZ adduct exhibits no color. In the presence of Chloride, Hg²ᶧ forms HgCL2 freeing up TPTZ which then binds the available Fe²ᶧ giving a very intense absorbance with a λmax~ 620nm. The assay is linear in the range 20 to 120 nmol Chloride/well with detection sensitivity ~0.4 mM chloride.

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What is the color of the chloride reagent?
The chloride reagent is supposed to be blue in color. The chloride reagent can be warmed to 37C and then mixed well to dissolve all precipitates. If there are still some precipitates, simply centrifuge and take the solution from the top.
Can EDTA interfere with the assay?
A metal chelator like EDTA might have effects on the reaction and hence should be avoided.
What is the assay principle?
The brief principle of this assay is as follows: TPTZ + Hg++ + Fe++: TPTZ-Hg binding is tighter, so Hg binds all TPTZ giving colorless complex. In presence of Cl- , HgCl2 is formed freeing up TPTZ which binds to Fe+2 giving deep blue color proportional to Cl- concentration.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
 BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Is the chloride reagent supposed to come in bright purple color? or should it be rather colorless?
The chloride reagent is bluish in colour to begin with. The final colour developed is going to be intense blue which can be detected at 620 nm.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration?
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.