Deproteinizing plasma?

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ntb103
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Deproteinizing plasma?

Post by ntb103 » Mon Jan 18, 2010 5:48 pm

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

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JackBean
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Post by JackBean » Mon Jan 18, 2010 7:33 pm

for glucose?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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biohazard
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Re: Deproteinizing plasma?

Post by biohazard » Tue Jan 19, 2010 7:35 am

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.

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Re: Deproteinizing plasma?

Post by ntb103 » Tue Jan 19, 2010 10:50 am

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

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JackBean
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Post by JackBean » Tue Jan 19, 2010 12:38 pm

He's talking about plasma, not plasma membrane! :lol: :twisted:

Sorry, I don't get, what you mean by "for glucose"
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

ntb103
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Re: Deproteinizing plasma?

Post by ntb103 » Tue Jan 19, 2010 2:15 pm

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?

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JackBean
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Post by JackBean » Tue Jan 19, 2010 2:40 pm

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.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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Re: Deproteinizing plasma?

Post by ntb103 » Tue Jan 19, 2010 5:24 pm

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

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JackBean
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Post by JackBean » Wed Jan 20, 2010 4:03 pm

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...
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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Post by bdastiarraga » Wed Nov 23, 2011 2:30 pm

have you found the protocol for blood glucose?

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Post by ChristopherC4rm34n » Tue Sep 09, 2014 6:15 pm

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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