This product includes 100 ul of 1000 x CMC dye. It is for measurement of 1000 samples using 96-well plates. It can also be used for determination of detergent CMC values using cuvettes.
The critical micelle concentration (CMC) value of a detergent depends on the solution compositions such as salt concentrations. The CMC assay kit is based on the principle that the detergent interacts with the fluorescence dye and enhances the fluorescence intensity at 465 nm (excitation at 360 nm). The kit can be used for measurement of the CMC values of known detergents in solutions with different salt concentrations that affect the CMC values. It can also be used to measure the CMC values of new detergents or to test if a new molecule is a detergent that should have a CMC value.
The CMC assay kit includes 100 µl of 1000 x CMC dye. It is for measurement of 1000 samples using 96-well plates. It can also be used for determination of detergent CMC values using cuvettes.
Concentration measurement
Detergent and lipids
Reference
1. Loreen R. Stromberg1 et al, Presentation matters: Impact of association of amphiphilic LPS with serum carrier proteins, on innate immune signaling, PLOS One, June 2018.
2. Kubicek-Sutherland J.Z. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. Toxins 11(4), 202 (2019).
ASSAY PROTOCOL
The following protocol is for assays using a standard black 96-well plate (Greiner 655076). Adjust the assay reagent volumes proportionally for measurements using cuvettes.
1. Prepare standard detergent solutions by a serial dilution of the detergent from 0.5 % to 0.0005% in water or a buffer. Set a control sample without detergent.
Note: This concentration range applies to most detergents. A different concentration range may be used for detergents with unusually high or low CMC values.
2. For each 100 samples, dilute 10 µl of the 1000 x CMC dye with 10 ml of water.
3. Mix 100 µl of the detergent solution with 100 µl of the 1 x dye and incubate the solution in the dark for 30 min.
4. Read the fluorescence at 465 nm (excitation at 360 nm).
DATA ANALYSIS
1. Calculate the ratio between the fluorescence of the samples with detergent and the control sample without detergent.
2. Plot the fluorescence ratio versus the detergent concentration. The transition point is the detergent CMC value.
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