Rabbit cells to grow bacteria in before immunization of rabbit for antibody prod
Posted 06 January 2011 - 01:11 PM
Since I've previously been helped here before and I'm not able to what I want to find/know online, I'm turning to you again!
It's kind of a long title, but essentially I'm looking for rabbit cells that I can use to grow up my bacteria. The genus/strain I'm working on is very uncommon in use and there are almost no antibodies out there in order to do much of my projected research. So I figure... I'll make my own. Since this is something we've done previously in the lab, I should have it done pretty quickly. The problem with the previous polyclonal rabbit antibody that we've made is that there's so much background! Here we grew up the bacteria in Vero cells, isolated them mostly from the Vero's (but still had a lot of debris, since the bacteria is really sensitive, so it's hard to seperate them from the cell debris still in there without killing them all!) and injected that into the rabbit. As a result, lots and lots of background.
Now I figure (and please if you guys have other suggestions, I'll be very gratefull as well) that when I can grow up my bacteria in Rabbit cells, I might not have that much background later on in my experiments. I just have no clue about Rabbit cell lines, except for the list that ATCC has.
So, please help!
What kind of Rabbit cell lines have you used to grow up bacteria or even virusses succesfully?
Or is there any kind of Rabbit cell line that you've used for a different set of experiments that you think might work well in this case?
Again, any help is grately appreciated!
Posted 11 January 2011 - 07:58 AM
Sounds like you want to produce a purified polyclonal antibody against some cell surface determinate of bacteria. You obtained ab that appears to be highly cross reactive.
If you want polyclonal that is specific to your bacterial antigen you may have to do affiinity chromatography.
Another option would be to make sure you have a purified antigen preparation or alternatively use mice and obtain the "clones" you require.
Maybe one of the forum members with experience in Mab/Poly production can jump in and help... with some suggestions..
Posted 13 January 2011 - 09:44 AM
Does that make it more clear?