If you're looking for simple procedures, you should homogenize the fish tissue in an adequate buffer (either using an ultra-sounds probe or a mechanical homogenization approach), centrifuge the debris and use the supernatant in the quantification kit.

This wont render a protein extract neither clean enough for further applications, nor 100% efficient in the extraction of protein.

Another (the most important) consideration to make is the choice of buffer. Standard protocol involves using PBS, which I recomend for its simplicity and compatibility with most quantification kits. However, some other buffers might be more efficient.

As to this point, if you want further help, it would be nice for you to say what protein quantification kit you have available... when extraction is made with the sole purpose of quantifying protein, it is better to design homogenization having the kit in consideration.

thanks alot for your time and attention

the available kits are total protein Biuret colorimetric method (coloremetric determination of serum total protein ) , iam giong to use these kits with tissue homogenate.

Dear Afaf,

No problem. The protocol here described should work fine:

- Sample (optimized amount for your case) + Liquid nitrogen + Mortar and Pestle = Powder

- Powder + Buffer (PBS, for your case - see below) + Sonication = Homogenate

-Homogenate + Centrifugation 10000 xg/15min (preferably, at 4ºC) = Supernatant

-Supernatant is your protein extract to use in Biuret assay.

PBS suitable to this experiment:

50 mM sodium dihydrogen phosphate ;

50 mM disodium hydrogen phosphate ;

1 mM EDTA;

1 mM DTT.

pH: 7 - 9

A reference for this protocol:

http://www.sciencedirect.com/science/article/pii/S1470160X15003659

Good luck

thanks alot for your reply and attention 

i'd like to ask what is the suitable  amount of muscle tissue to be homogenized??

and can i dispense the first step and put the tissue directly in PBS and homogenize it with the homogenizer??

I believe you have the solution for your issue. Now, I am researching fish physiology too. I also you the homogenate of muscle for AchE activity and BCA Protein assay. However 2 both kits do not mention about how to homogenate tissue. So, can you share me some your experience of this.

Thank you!

dear Van Tran,

detailed information are mentioned here, wish to help you.

all the best

Hello, I need to estimate total protein content in fish muscle using Lowry method and I want to know how to prepare the tissue homogenate for it. I Have got quite a few protocols and hence got confused. Can you share some of your experiences with me?

Thank you

Dear Chinmoyee Bhattacharyya,

I wish this attached file to be helpful for you