fitc labeling

[Gary Butler]

Lynne_A_WHITEHEAD at UMAIL.UMD.EDU (lw75) writes:

>Hi there!

>I am looking for a protocol for labeling proteins with FITC.  One source I
>found it in is checked out of the library.  I was wondering if any one new
>of a journal reference or  another source to find it in.  Also any
>information on how the stain labels the proteins (ex. charges, ect.)  would
>be appreciated!

>Thanks in advance!

>Lynne Whitehead
>U of Maryland - Micro Dept

This is off the top of my head, so double check it.  Mix FITC with 
protein in a pH 9.0 buffer, incubate a short time at RT, neutralize, 
separate FITC-protein (void)from free FITC with a small G25 column, 
centricon or amicon or dialysis.  For references you might try to find a
procedure in one of the following:  Murray Deutscher volume (180/182/185?) of
Methods in Enzymology on Protein Purification or Mishell & Shigii book
"Selected Methods in Cellular Immunology"  (WH Freeman Inc.), or perhaps 
the old Weir Immunology book.  Getting stuff labelled will be pretty easy.
Optimization of molar ratio may take some T&E. Good luck.  GB        
-- 
Gary H. Butler, Ph.D.
Coriell Institute for Medical Research
401 Haddon Ave.
Camden, NJ 08103

Tel: (609) 757-9716
Fax: (609) 964 0254 Attn: Butler
Email: butler at UMDNJ.edu