lab hints

Steve Holland holland at gasmac.dom.uab.edu
Mon Dec 13 16:57:07 EST 1993


In article <1993Dec13.150114.1 at earth.doane.edu>, dkatz at earth.doane.edu
wrote:
> 
> Since this is to be the first semester for me to teach immunology, I will
> probably be asking some questions of you tolerent immunologists this next
> semester.  Please bear with me.  I will be doing a lab with the course, and
> have come up with a couple of ideas.
> 1.  One of the standard lab experiments is to immunize and follow the course
> of the antibody production.  If we do this, it will have to be in mice, since
> we have not got the facility to handle larger animals.  Has anyone ever thought
> about getting serum samples from rather large puppies as they undergo their
> puppy shots ,and following the antibody titers this way?

An interesting idea.  the problem will be getting antigen to plate for
an ELISA.  I suppose you could go with an Ouchterloney, but an ELISA
is more current.  You might look into  donations from the company that
makes the vaccines.  They might also have some info on their results
with antibody levels developed after vaccination.

> 2.  Another technique that is often used is having students isolate antibodies
> from serum samples.  Off hand, does anyone know if it is very different to take
> antibodies out of colostrum or milk samples?

Well, the technique to draw the sample will be different.  An interesting
aspect would be to look at colostrum vs milk antibody levels, colostrum
is much higher.  Local dairy farmers might be able to give you milk
samples from just after vaccination to see what happens.  One practical
problem is eliminating the fat.  a spin in a refrigerated ultracentrifuge
works well.  there is also a product called lipo-clean that may work
on milk.  It is great for ascites.  Salivary antibody levels are another
source of antibodies.  More practical than stool antibody levels in humans
under the circumstances.  There was an article that described measuring
stool antibodies in mice - you can extract IgA from the pellets in mice.


> 3.  If anyone has any other ideas that are used in immuno. labs, I'd be very
> interested in speaking or emailing with you.  Budgets are limited, but I want
> to give my students the best experience an old bacterial geneticist can give in
> immunology.


How about skin transplants in mice across strains and in and out of 
offspring and F2's and the like?  Could get some cell lines responsive
to cytokines going and do some assays.  weHI 279 grow without cytokine
and are killed by interferon gamma.  Maybe some IL-1 assays using
thymocyte proliferation?  

best of luck with the course.
Please post some of your plans.  I would like to akeep notes on a low cost
immunology lab experience for future teaching I will do.

steve Holland



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