Quantifying Proteins, help required! - (Jan/11/2007 )
Hello all
Does anyone have any ideas on how to quantify a protein from wheat flour? The problem is, its difficult to extract on its own into a pure sample without losing any in the process, and i'm really interested in finding out exact amounts of it in wheat flour. I have a really good specific antibody for the protein but am unsure if i can quantify without purifying first?
Thanks for any help you may have.
If you need to quantify all proteins I would suggest Kjeldahl method and appropriate coefficient for wheat flour (look for some kind of food industry standards - they should have one for wheat).
thanks K.B, i re read my message and it isnt all that clear...
what i mean is, can i extract total protein, (and if so any ideas how to do this accurately) and then either use myy antibody to quantify my particular protein from the whole extract or any other method to pick out my protein, i know the exact size etc
What protein is that anyway?
YOu can quantify by doing a Western on your protein lysate ....
If you want to obtain exact # of ug, you should also run a std curve using the peptide htat was used to generate the Ab.
This method is reasonably accurate, although not perfectly quantitative. But you don't have any issues of protein loss and is quite generally done.
Does anyone have any ideas on how to quantify a protein from wheat flour? The problem is, its difficult to extract on its own into a pure sample without losing any in the process, and i'm really interested in finding out exact amounts of it in wheat flour. I have a really good specific antibody for the protein but am unsure if i can quantify without purifying first?
Thanks for any help you may have.