I want to measure the concentration of protein in given mucus samples , the problem is that during sample collection a lot of water may comes in yyour samples which makes your sample diluted and makes it also difficult to find a suitable blank. Do you have any recommendations ?
I wonder whether centrifugal filtration devices like Microcon-YM10 to concentrate your samples will be helpful!
Not sure if this is useful in the context of your samples, but if you can correlate the overall protein concentration to other components of mucus (i.e. glycoproteins, mucins) it might be possible to get a handle on it.
Navneet Ammal Kaidery,
I can understand that this may be helpful only for the problem of the dilution , however the problem of having a cut line for kDa passing to the lower compartments of the spin tube (eg.10 kDa) may affect my sample contents.
Peter Teriete
Actually, I didn't tell the whole story.
In fact, I want to measure specific Abs in fish mucus and I am thinking in the total mucosal protein or even Immunolobulins to correlate my ELISA reads out to.
Haitham,
It it should be possible to lyophilize the samples in previously tarred vials. Then you can measure the overall amount, take your sample up in an appropriate buffer, and run your ELISA in some and either a BCA assay on the other or see if there is an ELISA/Assay for a 'housekeeping' protein to correlated against. If you flash freeze in liquid nitrogen, I would expect very little impact on the proteins in the samples. I know of a neighboring lab that analyzed monkey feces this way for various microbial and viral content. It was always interesting coming to the freeze-dryer and seeing flasks full of monkey s***.
Peter Teriete. this is a very good idea I think it worth atrial
Thanks for kind help
Total protein conc. can be measured using Lowry or Bradford assays. They are very simple and fast.
Yes sure IIgin Paker, it is very simple and easy , but the problem is that when I want to dilute BSA in buffer "X" , and my mucus sample is not diluted in the same buffer, hence it is not a convenient way and I can not even dilute it because I need to measure specific Ab in the mucus which is hardly detected, anyway thanks for your answer
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