I want to measure the diameter of High density lipoprotein (HDL) in Tris buffer by Dynamic Light Scattering (DLS). However, these proteins are binding together. Could anyone help me, how can we avoid HDL aggregation?
Thank you in advance,
Maybe you should try to run simulation increasing the salinity percentage in a system.
Varying the ionic strength could be helpful. If the aggregation is caused by salt bridges, increasing the ionic strength should help. If the aggregation is caused by hydrophobic interactions, decreasing the salt concentration, and possibly adding a little bit of a mild detergent, could help.
Hi Nam, you can try adding delipidated serum to the assay to prevent protein-protein interactions.
Hi Steingrimur, I think adding serum would cause a problem with the DLS measurement because of the proteins it contains, especially the high concentration of albumin.
Thank you all guys for your information.
Hi Adam B Shapiro, I agree with you that the problem may cause by the interaction of the hydrophobic core ( triglycerie structural) of HDL because I have tried to filter these protein by Miliporous Syringe Filter and it is not effective.
As your advise, Could I use sodium dodecyl sulfate (SDS) and Tween 20?
Thank you for your kind of feedback.
I wold not recommend using SDS because it is denaturing to proteins, although I don't know whether this is true for HDL. Non-ionic detergents like Tween-20, Triton X-100, and Brij-35 would be preferable. Use a high-quality source (I recommend Surfact-Amps from Thermo-Fisher Scientific) and keep the concentration below the critical micellar concentration to avoid having micelles, which can show up in DLS. I suggest trying a concentration of 0.005%.
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