i am trying to prepare 8% casein in pbs for peritonial recrutment of neutrophils but i am facing problem in dissolving casein in pbs.
kindly suggest me the posible pitfalls as i am doing it for first time.
Daniel
Dissolving Caesin can be challenging due to its poor solubility in neutral pH and aqueous solutions. Casein is a milk protein that dissolves more readily in acidic conditions or with the help of heat and stirring. You can also add NaOH (that’s correct) dropwise to aid it in solubilization.
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We Pre-soak the casein in a small volume of PBS overnight at 4°C before heating to aid dissolution.
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Daniel You can also use Chelators like sodium citrate or EDTA can break down the micellar structure of casein by binding calcium ions, making it soluble. Dissolve the chelator in distilled water first, then gradually add casein while stirring. In case detergent doesnt interfere with your experiments, you can also add Tween 20 or Triton X 100
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