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in what technique can i use to detect specific proteins


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#1 liika

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Posted 04 March 2014 - 10:28 AM

 inside a bacterial cell? i mean, assuming i have some bacteria, and i want to check if it's synthesizing a specific protein, then what method should i use?

 

thanks!



#2 pito

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Posted 04 March 2014 - 11:47 AM

There are many techniques.

 

What about a qPCR? Do you know the gene?


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 liika

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Posted 04 March 2014 - 12:46 PM

There are many techniques.

 

What about a qPCR? Do you know the gene?

i do, but i want to check specifically the product itself-the protein. not the gene...



#4 pito

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Posted 04 March 2014 - 12:48 PM

well qPCR checks wether the DNA is translated... so it also says something about the protein.

Other ways are , for example, co-immunoprecipitation, Y2H, affinity tagging, GFP tagged proteins...


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#5 liika

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Posted 04 March 2014 - 12:50 PM

ok, thanks a lot



#6 mdfenko

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Posted 05 March 2014 - 04:43 AM

does the bacteria have to remain intact or is the protein exuded into the medium?

 

if it doesn't have to remain intact or if it is exuded then you can perform a western blot or elisa with the extract or medium.

 

if the bacteria has to remain intact and the protein is not exuded:

 

is the protein an enzyme?

 

if so, then you may be able to use a chromogenic or fluorescent substrate or cofactor if it can enter the cell.


Edited by mdfenko, 05 March 2014 - 04:47 AM.

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