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method to separate and quantify plasma proteins - please help! (Jun/12/2008 )

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anyone could tell me a nice and accurate method or device, which could separate and quantify each protein concentration within plasma solution please blink.gif ? Thank you very much

-Essell-

QUOTE (Essell @ Jun 12 2008, 07:43 AM)
anyone could tell me a nice and accurate method or device, which could separate and quantify each protein concentration within plasma solution please blink.gif ? Thank you very much

If I understand your question correctly:

Plasma solution has about 100,000 different proteins, ranging from milligram to famtograms in conc. There is no way to identifyeach (unknown) protein.

You can of course do MS/MS and look at peaks and identify many proteins by tryptic digest pattern and edman sequencing, but not all. You need some way to separate the plasma (size exclusion etc) and still if the protein does not exist in the database, it will not be identified.

-cellcounter-

QUOTE (cellcounter @ Jun 12 2008, 11:44 PM)
QUOTE (Essell @ Jun 12 2008, 07:43 AM)
anyone could tell me a nice and accurate method or device, which could separate and quantify each protein concentration within plasma solution please blink.gif ? Thank you very much

If I understand your question correctly:

Plasma solution has about 100,000 different proteins, ranging from milligram to famtograms in conc. There is no way to identifyeach (unknown) protein.

You can of course do MS/MS and look at peaks and identify many proteins by tryptic digest pattern and edman sequencing, but not all. You need some way to separate the plasma (size exclusion etc) and still if the protein does not exist in the database, it will not be identified.


thank u mate! can i also separate them and quantify each of them, possibly?

-Essell-

QUOTE (Essell @ Jun 13 2008, 02:44 AM)
QUOTE (cellcounter @ Jun 12 2008, 11:44 PM)
QUOTE (Essell @ Jun 12 2008, 07:43 AM)
anyone could tell me a nice and accurate method or device, which could separate and quantify each protein concentration within plasma solution please blink.gif ? Thank you very much

If I understand your question correctly:

Plasma solution has about 100,000 different proteins, ranging from milligram to famtograms in conc. There is no way to identifyeach (unknown) protein.

You can of course do MS/MS and look at peaks and identify many proteins by tryptic digest pattern and edman sequencing, but not all. You need some way to separate the plasma (size exclusion etc) and still if the protein does not exist in the database, it will not be identified.


thank u mate! can i also separate them and quantify each of them, possibly?


not with simple methods;, try 2D-GE, identify each polypeptide, as suggested by Ms/Ms, and semi-quantify by staining

-The Bearer-

QUOTE (The Bearer @ Jun 13 2008, 02:10 PM)
QUOTE (Essell @ Jun 13 2008, 02:44 AM)
QUOTE (cellcounter @ Jun 12 2008, 11:44 PM)
QUOTE (Essell @ Jun 12 2008, 07:43 AM)
anyone could tell me a nice and accurate method or device, which could separate and quantify each protein concentration within plasma solution please blink.gif ? Thank you very much

If I understand your question correctly:

Plasma solution has about 100,000 different proteins, ranging from milligram to famtograms in conc. There is no way to identifyeach (unknown) protein.

You can of course do MS/MS and look at peaks and identify many proteins by tryptic digest pattern and edman sequencing, but not all. You need some way to separate the plasma (size exclusion etc) and still if the protein does not exist in the database, it will not be identified.


thank u mate! can i also separate them and quantify each of them, possibly?


not with simple methods;, try 2D-GE, identify each polypeptide, as suggested by Ms/Ms, and semi-quantify by staining


thanks a lot!

-Essell-

If you're going to do 2D gels, consider pretreatment with anti-albumin, so you can see the lower-abundance proteins...

-swanny-

QUOTE (swanny @ Jun 15 2008, 04:40 PM)
If you're going to do 2D gels, consider pretreatment with anti-albumin, so you can see the lower-abundance proteins...


interesting, swanny...but how does it work? and for what staining method? or Ms/Ms spectra?

-The Bearer-

QUOTE (swanny @ Jun 16 2008, 12:40 AM)
If you're going to do 2D gels, consider pretreatment with anti-albumin, so you can see the lower-abundance proteins...


do u mind to give more details? like why?

-Essell-

because you can load more proteins without albumin.

-genehunter-1-

QUOTE (genehunter-1 @ Jun 16 2008, 03:11 PM)
because you can load more proteins without albumin.


see. great! thank u guys.

-Essell-

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