In article <D1vtry.vJ at ncifcrf.gov>, reming at ncifcrf.gov (Mary Remington) says:
>> We are currently using western blots to determine expression of a protein.
> Our antibody has not been impressive so far. It has been suggested that we
> use immunoprecipitation instead of the western technique. Can immunoprecipitation
> be used to compare amount of protein expression or is the western the
> better way to go? Thanks for your input. Mary
Mary,
Both techniques have their pros and cons. To quantitate a radioactive
signal in a western, you must work within the linear range of your system, i.e., run several
different amounts of your purified protein on your gel, do western, and scan the autoradiogram
with a densitometer or similar device. If the area of integration is linearly proportional to your
mass, then you can use the western to do what you want.
In immunoprecipitation is used, then you must be very careful to process all
samples equally. You can easily loose protein during the washing stages. Otherwise,
this technique is another reliable method. Hope this helps.
Jim Bassuk
BioStructure
Univ. Washington
