Hi everyone
I want ant to use ELISA to quantify tnf a ,il-1b and inf g in Half brain of mice. I was wondering if any one has done this exactly before and can tell me which concentration of total protein he used ?
Dear read the protocol provided by the manufacturer of the kit you use.
Hi Esraa,
Your total protein concentrations will vary from sample to sample. First, it is important to know the range of your standards for your ELISA kit, based on that range you can standardize the total protein concentraiton you want to analyze with that ELISA. Next, you need to determine the total protein concentration for each sample you have collected. One method that is quick and relatively inexpensive for determining total proteins would be a Pierce BCA Protein assay kit. The results of that assay will give you a total protein cocentration per brain sample. From there, you can titrate your individual brain samples to your desired standardized concentration based on the limitations of your ELISA kit. Then simply following the directions of your kit and analyzing your results. Hope this helps!
G. Andrew Pratt III Andrew Pratt III
thank you for reply, was really helpful.
The kit has a determination limit up to 1000 pico gram. I read in one article that they used 100 ug of total protein , that mean the sample concentration was 1 mg/ml . I was wondering if any one did exactly the same thing so I would be sure.