ELISA Problem since change of antibody supplier? - (Apr/24/2008 )

Hello All

This is my first post on here so hopefully you can help me out!

We've been running an optimised, indirect ELISA for a few months now which has been working perfectly.
Unfortunately the company which makes the primary antibody stopped selling it. After a minor panic we found an alternative company who can also supply the antibody.

However, the new antibody just doesn't work. There is no signal over background at all. I did a side by side comparison with both antibodies on the same using the same set of lysates & secondary Ab, & the difference between the two is like night and day.

Looking at the data sheets the antibodies are definitely raised from the same source. The only difference is that they are supplied in slightly different buffers and protein concentrations.
I did one experiment titrating lysate & primary concentrations but still couldn't get a signal.

Has anyone had a similar problem before? I've contacted the company but thus far haven't had a reply.

Many Thanks for any help
P

Hello All

This is my first post on here so hopefully you can help me out!

We've been running an optimised, indirect ELISA for a few months now which has been working perfectly.
Unfortunately the company which makes the primary antibody stopped selling it. After a minor panic we found an alternative company who can also supply the antibody.

However, the new antibody just doesn't work. There is no signal over background at all. I did a side by side comparison with both antibodies on the same using the same set of lysates & secondary Ab, & the difference between the two is like night and day.

Looking at the data sheets the antibodies are definitely raised from the same source. The only difference is that they are supplied in slightly different buffers and protein concentrations.
I did one experiment titrating lysate & primary concentrations but still couldn't get a signal.

Has anyone had a similar problem before? I've contacted the company but thus far haven't had a reply.

Many Thanks for any help
P