Biotin block with eggs and milk

From:Phyllis Davie <pdavie@phenopath.com>

Hi all,
I'm reposting this since I think my new e-mail program inserted an
attachment to my previous posting (probably my signature).  Sorry if this is
a duplicate.


Dr. Karimi,
  Here is the eggs and milk recipe we are using, with good results.  Hope
you find it useful.


Phyllis Davie
PhenoPath Laboratories--Seattle, WA
pdavie@phenopath.com



1.1.    Biotin Blocking--Version 2 Eggs and Milk

1.1.1. Reagents

1.1.1.1. Egg Substitutes:
Commercial egg substitutes such as "Second Nature Real Egg Substitute"
(Product Ingredients: A Pasteurized Blend of 98% Egg Whites Containing Less
than 1% of the Following: Corn Oil*, Polysorbate 60*, Sodium Stearoyl
Lactylate*, Salt, Xanthan Gum, Guar Gum, Beta Carotene (Color), Onion
Powder)  Purchased from any grocery store.
Storage:  refrigerated (2-8C);  do not freeze
Shelf Life:  use by expiration date on package
PREPARATION:  
In the container to be used, mix 200ml diH2O   and  75ml egg substitute.
Storage of working solution: refrigerated (2-8C);  do not freeze.
Shelf Life of working solution: one day

1.1.1.2. Milk, fat-free:
Purchased from any grocery store.
Storage:  refrigerated (2-8C);  do not freeze
Shelf Life:  use by expiration date on package
PREPARATION:  
Use as supplied
Storage of working solution:  refrigerated (2-8C);  do not freeze.
Shelf Life of working solution:  one day


1.1.2. Procedure:    Biotin Blocking--Version 2Eggs and Milk
1.1.2.1. Incubate slides in the egg solution, in racks, for 15 minutes at
room temperature. 
1.1.2.2. Pick up all the slides and wash in 2 changes of di H2O.
NOTE:  Do NOT wash in buffer, as buffer will precipitate the egg.
1.1.2.3. Incubate slides in the milk, in racks, for 15 minutes at room
temperature. 
1.1.2.4. Pick up all the slides and wash in 2 changes of di H2O.
1.1.2.5. Continue with the desired procedure at the antibody application
step. 


1.2. References:  Both Versions
Matsumoto Y.  Simultaneous Inhibition of Endogenous Avidin-Binding Activity
and Peroxidase Applicable for the Avidin-Binding System Using Monoclonal
Antibodies.  Histochem 1985; 83:325.

Wood GS and Warnke R.  Suppression of Endogenous Avidin-Binding Activity in
Tissues and its Relevance to Biotin-Avidin Detection System.  J. Histochem
Cytochem 1981; 29:1196.



on 5/7/01 7:02 AM, shirin karimi at shirinkarimi_1@yahoo.com wrote:

> Hi,
> How can I use egg as a biotin blocker.
> The commertial biotin blockers are not avaible for me.
> Thank you,
> 
> Shirin Karimi,M.D.,A.C.P.
> 
> __________________________________________________
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