western blot

I am new to western blot and experienced that when i store my lysate at -20, and freeze thaw it , and run the gel, it works wel and gud band seen after developmentl, but after that if i repeat the same sample again after freeze thawing, no band was seen after development. I have read that protein lysate shud be stored at -20 or -80 and in aliquotes.
So, can by storing lysate at -80 and in aliquotes, if i run the sample sample and can get gud band.

I have not made aliquotes as i my lysate was suspended in very less buffer and also as i thought repeated freezing thawing of same lysate will not have problem if i am freeze-thawing it just two/three times.

can anybody can tell me how i can get gud band everytime by same sample once i freeze-thaw it.

thanks in advance.