What is the sensitivity of dot blot compared to ELISA, and what antibody concent - (Feb/06/2007 )

Hi,

I'm going to do dot blots but I have no idea what's a good starting dilution to use of my various antibody sources.

In some cases for the pure antibodies, a good ELISA titer is given. Should I use a more diluted solution for dot blot than what is suggested for ELISA?

In one case the potency of a purified antibody is only given as in relation to that of cell culture supernatant but I have no idea what dilution one would generally use of cell culture supernatant, or of ascites fluid. What antibody concentrations can I expect from cell culture supernatants and ascites fluids?

Help, please!

Mab conc in cell culture super is varied between 10-100microgram\ml depends on cultivation technic and in ascite fluid near 1-3 mg \ml. I think you may start for dot with Mab (cell culture) 1:50 dilution and for ascites start from 1:1000.