Hello everyone! I am currently working on the expression and purification of an antimicrobial peptide that experiences reduced activity after each freeze-thaw cycle. To improve its stability, I need to incorporate sucrose. However, the presence of sucrose in the dialysis buffer is causing issues with determining the peptide concentration, as it results in a high purple color. Does anyone have any suggestions on how I can address this problem?
Tlehema Umbayda
One option is the use of colorimetric methods that specifically target peptide quantification. diluting the sample before measurement might help reduce the color intensity of sucrose. Another approach could involve performing dialysis using a sucrose-free buffer after incorporating the peptide, allowing me to assess concentration without interference. Creating a standard curve that accounts for sucrose concentrations could also aid in correcting absorbance readings. Use other stabilizers that do not interfere with concentration measurements, such as trehalose or glycerol. Last but not least, if the peptide possesses distinct UV absorbance characteristics, using UV spectroscopy for direct quantification could provide a clearer assessment.
Hi, everyone on this platform
To address this issue, I am considering several strategies.
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