Immunobloting of Albumin...

[Roman Fried]

Paulo Barros Alexandrino (pbalexan at fc1.fc.up.pt) wrote:
: : Dear All
: : I would like to know what kind of blocking agent would be advisable
: : to employ for immunological probing of albumin after transfering the
: : protein to a nitrocellulose matrix. Use of gelatin or non-fat milk
: : is yelding a too undesirable background.
: : Thanking in advance any answers...
: : Paulo Alexandrino
: : University of Porto, PORTUGAL
: : E-mail: pbalexan at fc1.fc.up.pt



Dear Paulo

In our lab we use the following protocol

1. incubate the nc membrane 1 h in tween-tbs (tris 20mM, NaCl 0.5M,
pH 7.5 and 0.3% tween-20

2. 1. antibody over night (in tbs-tween)

3. 2 x 10 min tween-tbs

4. 2. antibody 1h (in tbs-tween)

5. 2 x 10 min tbs (no tween!)

6. detection with chloronaphtol 

7. after detection you can do an overall staining
using colloidal gold solution

i hope this helps

regards 

roman

Roman Fried
Institut fuer Klinische Chemie
Universitaetsspital Zuerich
8091 Zuerich

mail rmf at ikc.unizh.ch