Hey Tiffy!, unfortunately it seems that it may be your antibodies. Which species did you use? and which types (specificity)?
reduced and denatured rabbit IgG heavy chain band runs at ~ 50 kDa
close to your size....
also 25 kDa may be the other antibody fragment...
please post the species info and your protein of interest and we may help more
Originally Posted by Tiffy
is it right that under optimal parameters I got only one band after IP and Western Blot? Today I did my first IP and I got 2 bands but not my band of desire
The bands were at ~ 45-50 kDa and ~ 20 kDa. Is this the destroyed fishing-ab (heavy and light chain)?
It would be very kind if someone could answer my question.