Blocking with Milk

[Ri-Ya Yang/SBR4/4-9980]

I usually use 10% milk in TTBS to block Immobilon-P PVDF membranes 
(Millipore) at room temperature for 10min. The trick is you should 
prepare milk solution immediately before use.

Ri-Yao
Dept.Mol.Exp.Med
The Scripps Res.Inst.


On 22 Sep 1995, Sucking Gippy wrote:

> Just a general question with regards to blocking membranes before 
> probing. I generally use a non-fat milk solution in TTBS.
> Question: What are ideal dilutions to use? i.e. I've tried everything from
> 	  2%--> 10% Comments?
> 
> 	  What is an ideal time period? Most "gaps" on the membrane (usually
> 	  nitrocellulose) should be blocked immediately upon addition of 
> 	blocking agent...it would be those so called "deep down pores/holes"
> 	 where a time period for blocking would be effective. I guess the 
> 	 membrane pore size would also have an effect on the time period
> 	 required..right? Generally I use 1 hour at room temp with the 2% milk
> 
> 
>