which company can raise good antibodies? - raising antibodies using the native protein (Oct/11/2006 )
hi, guys,
my lab is working with a protein. We need to see the localization of the this endogeneous protein. however, we cannot find a good antibody in the market even we submitted a peptidtes from c or N terminus for raising antibidies. So, I would like to ask, do anyone have experience about which companies can raise good antibodies? I would like to use the whole protein to raise the antibody or using a fragment around 200 aa. How long will it take to raise an antibody like this?
thanks
my lab is working with a protein. We need to see the localization of the this endogeneous protein. however, we cannot find a good antibody in the market even we submitted a peptidtes from c or N terminus for raising antibidies. So, I would like to ask, do anyone have experience about which companies can raise good antibodies? I would like to use the whole protein to raise the antibody or using a fragment around 200 aa. How long will it take to raise an antibody like this?
thanks
If you use the whole protein or the more aa the better, you will get better antibody. Which companies have you tried? Do you know what their immunization schedules were like? Did you purify the serum you received?
At the begining, We used the in house facility in the Univeristy. However, none of them are nice. Then, we submit the N terminus and C terminus to Invitrogen (As I remember), they have several polyclonal antibodies, two of them can identify the protein properly from western blot. But still have a nonspecific band. Then, we tried to purify the antibodies, the signal of the target becomes weaken, and another 2 nonspecific bands appear!!!! howcome.
I need antibodies for immunohistochemistry.
If the signal was weakened, it could either mean that non-specific antibody got caught up in your purification somehow, or that you just need to use a higher concentration of it. Have you tried those antibodies in IHC yet? They may work just fine. I have used Strategic Biosolutions and Lifespan Biosciences, and I was happy with both. Try your antibodies in IHC first, just in case they work fine so you don't have to spend more money!