For isolation of PBMC, at what temperature we should keep sucrose solution.
Which method is better for PBMC isolation? Ficoll DGC method or Sucrose DGC method?
Uncontaminated Ficoll solution can be stored at 4°C indefinitely.
For PBMC isolation, I recommend Ficol. Unlike sucrose, solutions of Ficoll have relatively low osmolality. Despite this, Ficoll's density in aqueous solutions is comparable to that of sucrose. Because of the high molecular weight and low content of dialyzable material, Ficoll has a much lower permeability towards cell membranes than sucrose.
It can vary depending on the experimental requirements and the nature of the sample being studied. However, a commonly used temperature for sucrose gradient centrifugation is 4 degrees Celsius (4°C) or refrigeration temperature.
Keeping the sucrose solution at 4°C helps maintain the stability and integrity of the sample during the centrifugation process. It helps reduce the degradation or denaturation of biomolecules, especially proteins and nucleic acids, that could occur at higher temperatures. Cooling the sucrose solution also helps prevent the diffusion or mixing of the gradient layers, allowing for better separation and resolution of the sample components.
Hope it helps.credit AI
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