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Western Blot Analysis - Technique for Protein determination (Jul/18/2006 )

Hi all,

I'm a pharmacy student from India looking for some features concerning Western Blotting. I would like to know abt. the types of samples which can be used in this technique. And also is this is the only way to determine protein samples found in the given sample.

-Arulnathan-

QUOTE (Arulnathan @ Jul 18 2006, 05:27 PM)
Hi all,

I'm a pharmacy student from India looking for some features concerning Western Blotting. I would like to know abt. the types of samples which can be used in this technique. And also is this is the only way to determine protein samples found in the given sample.

Check http://en.wikipedia.org/wiki/Western_blot for information about the procedure. YOu ca of course also enter the search term "Western Blot" on the protocol-online.org site.
What do you mean by type of samples? You can basically apply this method to detect any protein (if you have an antibody against it) in a given sample containing a mixture of proteins .

-Jou-

Hi Jou,

thanks. I asked abt. the types of samples.....like whether it could be blood or some other tissue samples.
I got some tips from wikipedia.

-Arulnathan-

Arulnathan, there is a hindrance to running something like blood or tissue directly on a gel. you can certainly use the Western technique, but first you need to do a total or partial protein preparation of the tissue / blood/ etc you want to test. This allows you to remove or inhibit proteases present in your rough sample, and also to get a good homogeneous sample to work with.

does this help?

-aimikins-

Thanks aimikins, Yes. I proceed with my plan...as you mentioned ..I prepared the sample to be estimated but
still I facing some probs with this step itself.

-Arulnathan-

hi

other ways to detect proteins is ELISA or dot blot or immunoblots

of course first u do need a good protein extracted from the tissues


all the best!

-phytoviridae-

About the type of samples, I do WB using human serum (of course first you obtain the blood, centrifugate and collect the serum). It´s not usual because I think that most proteins in serum are determined by ELISA or similar, but when you don´t have the ELISA, WB is quite useful, at least before preparing the ELISA, only if you want to know if a specific protein is present in your sample. Well, that´s my opinion, but I´m not an expert.

-pumuki-