(>Ron Tate) Re: Immunoscreening cDNA expression library

Hiranya Roychowdhury hroychow at NMSU.EDU
Thu Oct 30 18:27:46 EST 1997


At 11:24 AM 10/30/97 -0800, Snyder Research Foundation Staff wrote:
>Dear Group,
>	I am thinking about screening a cDNA library with an antibody we 
>have for a protein which cross reacts with the antibody.  I would like to 
>hear from people who have actually done such screening so that I know some 
>of the pitfalls I may face before they hit me in the face.

The antiserum needs to be "pre-absorbed" against the host bacterial lysate
to overcome the background cross-reactivity on the lifts. If you need a
protocol for that, email me.

  Also, does 
>anybody have any recommendations about commercially available kits, and 
>thoughts about 125I versus Alk. Phos'tase (the two kits I have found are 
>both alk. phos'tase based).  
>

125I is messy and is better avoided. AP protocols work fine and you do not
need any x-ray film for it. I am not sure if you really need a kit for it
though. You can purchase the AP-conjugated seconfdary ab, NBT, BCIP from any
source and the salts needed for the buffers are there in your lab.


Dr. Hiranya Sankar Roychowdhury
Plant Genetic Engineering Lab.
New Mexico State University
Las Cruces, NM 88003
Ph. (505) 646-5785
hroychow at nmsu.edu




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