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    • #12631
      ntb103
      Participant

      Does anyone have a step by step protocol for deproteinizing plasma for glucose with barium hydroxide and sulfate?

    • #96806
      JackBean
      Participant

      for glucose?

    • #96809
      biohazard
      Participant

      If you deproteinize plasma theres nothing much left 🙁

      All you got left is some glucose, ions, non-protein hormons and such… and a lot of water.

    • #96814
      ntb103
      Participant

      Yes, for glucose!! Does anyone have a protocol?

    • #96817
      JackBean
      Participant

      He’s talking about plasma, not plasma membrane! 😆 😈

      Sorry, I don’t get, what you mean by "for glucose"

    • #96820
      ntb103
      Participant

      Yes, blood plasma. I need to infuse radiolabelled glucose in vivo and measure hepatic glucose production. I am just not sure the best way to deproteinize the blood to get the infused glucose vs. blood glucose values?

    • #96823
      JackBean
      Participant

      What scale do you need? How many samples? Do you need it online?

      What about to try desalting column of gel filtration?

      EDIT
      or you can try precipitation, either by ammonium sulphate, organic solution, or probably most appropriate for you by heat.

    • #96826
      ntb103
      Participant

      Working with 10 microL of plasma per sample and about 10 samples per run. I am not sure what you mean by online?

    • #96867
      JackBean
      Participant

      By online I meant something like taking blood from someone,measure and probably give back…

      Well, with 10 ul, you can use some spin-filter or the precipitation…

    • #108336
      bdastiarraga
      Participant

      have you found the protocol for blood glucose?

    • #115425
      ChristopherC4rm34n
      Participant

      Hello! Here is a protocol currently used by a core facility for the deproteinization of small volumes of mouse plasma for measurement of plasma radiolabeled glucose:

      1) Add 10 uL of plasma into 20 uL of dd water.
      2) Add 25 uL of BaOH (0.3N from Sigma B4059) and then add 25 uL of ZnSO4 (0.3N from Sigma Z2876).
      3) Vortex and spin at 12,000 rpm for 10 minutes.
      4) Collect 25 uL in duplicate and then add to 3-5 mL of scintillation fluid for counting.

      I hope this helps someone even though I’m responding to a 3-year old question. If anyone finds this helpful please let me know!

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