3 replies to this topic

[IPI]

Hi There,

Back in August, I asked the following question:

As a first part of making a mouse monoclonal antibody against a murine cytokine by hybridoma cells, I was wondering:

Can an ELISA standard like mouse IFN-gamma (found in a commercial ELISA kit), mixed with an adjuvant (e.g. TiterMax) be injected to mice in order to induce the B-lymphocytes to generate anti-IFN-gamma antibody? Does this sound theoretically as a plausible approach?

And one of the contributors answered: Sure its plausible.  Since monoclonals bind to the standards clearly the standards are antigenic.  Since it is an endogenous protein and relatively small it might be beneficial to conjugate it to something like KLH to stimulate a strong response.  

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Now I have another question related to that topic: Considering the fact that most companies use rats as their host for producing mouse monoclonal antibodies, would it be practical to use mice such as BALB/c for this purpose, instead? I am considering this because we cannot have rats in our facility. By the way, I am going to use KLH with my antigen.

Edited by IPI, 17 November 2012 - 01:16 AM.

[Curtis]

yes, you can inject to them and get antibody, but you need anti-mouse secondary to detect them, not anti-rat.

[leelee]

No you can't, not possible.

You can't get a mouse to make antibody to one of it's own proteins. It must be an immunogen, which a self protein is not.

[Curtis]

i just read the question, not the protein type,sorry