western for ZO-1

Jes Kuruvilla jkuruvil at lamar.colostate.edu
Thu Jun 21 17:28:29 EST 2001


Hi Sid,

Do a 4-16% gradient gel available from Bio rad or use a 4%
stacking and 10% running gel. to separate all your proteins.
The use the entire gel including the stacking in a submerged
transfer apparatus (biorad) . This is the best way to
transfer a large protein. We use it for CBP(2441)
aminoacids.

proceed after transfer onto PVDF or Nitrocellulose and then
using a common western blotting scheme used for regular
proteins detect with a specific antibody.

Jes

Siddhartha S Mitra wrote:
> 
> can anyone give me the protocol for doing western blots for high
> molecular weight proteins (~220Kd) specifically ZO-1
> 
> --
> Siddhartha S Mitra
> Cooke 615
> Biological Sciences
> SUNY at Buffalo
> Buffalo NY14260
> 
> Tel:   (Lab)716 645-3323
>  (Res)716 837 2507
> 
> Email: ssmitra at acsu.buffalo.edu




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