as per my experience band does not come. i tried same like you but not cytochrome c i used PGC1 A antibody

By "rabbit secondary antibody" do you mean "anti-rabbit secondary antibody"? This will not work. Switch your secondary antibody.

And do go and read a little on western blot theory... It will save you a lot of time and money.

You can start (but don't stop) here

http://en.wikipedia.org/wiki/Western_blot#Two_steps

You could use protein A with a horseradish peroxidase tag. Protein A binds to Fc portions, witohut discrimination of antibody specificity, or animal origin. Is your cytochrome c very diluted? usually you can see the red band on your SDS-PAGE or IEF while running.

thanks a lot for your suggestions

Definitely you can use protein A with a horseradish peroxidase tag, but I fear much of the amplification effect will be lost. Also, will it actually be cheaper then commercial anti-rabbit-HRP Ab?

i am agree with yoram Gerchman , i have tried in my lab also surely there will be difference of amplication factor, so for better result you can go for anti-goat secondary antibody. Gud luck

U will never get a band on your western blot...... switch to anti goat secondary antibody,,,,, good luck

You have to use anti-goat secondary antibody if your primary ab is originated from goat.

Secondary antibodies are not as expensive as primary antibodies. You can buy good anti-goat IgG tagged with peroxydase to well known manufacturers like Sigma-Aldrich or R&D. It's better to use secondary antibodies to keep the aplification effect, especially if the concentration of your sample is weak.

You will most likely not get any bands if you try to use the rabbit, and even if you do, how could you determine what is real versus artifact? You need to purchase the appropriate secondary antibody. As Pierre-Marie says, they are very inexpensive and will ensure that you results come out correctly.

Protein-A-HRP is a good alternative but if I am not wrong the biding affinity is much lower than commercially available rabbit or donkey anti-goat-IgG-HRP. However, you may actually get a band, or even multiple bands, by using anti-rabbit-IgG-HRP. You may even see a band with the right moleculkar weight, which I have seen before, due to non-specific binding. As a cautious scientist like you I will suggest you no to bet on that.