Hello Researchers,
I am trying certain new antibodies for first time in our lab for cell culture experimental samples (N2A mouse neuroblastoma cells). Due to low protein concentration in cell culture samples, i do not think it will be wise to use those samples for standardization. So, can i use mice brain tissues to standardize antibody concentration for western's. Your answers are greatly appreciated. Thanks !
Hi Richard,
Are you lysing the cells and tissue to use the protein in a Western blot? There is more information required before a recommendation can be made. Are you sure the tissue also expresses the protein/s of interest? If so, you could certainly use tissue lysate to check that your antibody is recognised at the expected band, but you would still need to optimise for concentration with the cell lysate, since it may be very different.
Gae.
Thanks a lot Gae for your input. Yes, both the tissue and the cells express the proteins of my interest.