[Histonet] New rabbit antibody... but what do I use as negative (isotype) control?

[Gayle Callis]

Rabbit IgG will work - you can purchase from Jackson.  This is a 
polyclonal, and if there is no isotype listed, whole IgG will suffice.

The concentration of the IgG should match the concentration of your primary 
antibody, and calculate it in terms of ug/ml - a little math. Rather than 
rely on some dilution, you will dilute the Rb IgG, but its original 
concentration in mg/ml may be much higher than the primary antibody's 
mg/ml, meaning you might be using a different dilution for Rb IgG than you 
use for the rabbit primary antibody.

You could use rabbit serum, but it should be heat inactivated - if the 
polyclonal is affinity purified and the pooled serum is not, you may get 
more nonspecific binding with serum.  Rabbit IgG would be my choice.


t 08:08 AM 5/11/2005, you wrote:
>Hi everyone,
>
>I have a new antibody that I was given.  It is a rabbit polyclonal 
>anti-mouse antibody.  Usually I use a specific isotype control, but I do 
>not have info on how this antibody was made other than the epitope(s) that 
>it should adhere to...  but I want a negative control and was wondering if 
>I can just use regular rabbit serum (diluted) as a negative seeing that it 
>should contain the isotype IgG I am using??? I think I am a bit confused 
>here.... I need assistance.
>
>Can I use rabbit serum as a negative control?
>What dilution do I use if that is the case?
>Anything else I should know before I proceed?
>
>Thank you all!!
>
>
>
>
>---------------------------------
>Discover Yahoo!
>  Use Yahoo! to plan a weekend, have fun online & more.  Check it out!
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)