Q: Fluorography of SDS-PAGE, need protocol

Jonathan B. Marder MARDER at agri.huji.ac.il
Thu Feb 23 02:58:54 EST 1995


If  you after quality rather than speed, I would recommend the old and tested
PPO/DMSO method (Bonner WM, Laskey RA, 1974, Eur. J. Biochem. 76, 83-88)
This involves putting your gel through:-
    2 X 30 minutes DMSO
    2-3 hours in 22% PPO dissolved in DMSO
    60 minutes wash in running tap water (PPO precipitates in gel).
    Dry

a.devitt at bham.ac.uk (Chutney) wrote:
>In article <3hgg2v$qkm at usenet.INS.CWRU.Edu>, zxy5 at po.CWRU.Edu (Zafer
>Yildirim) wrote:
>
>> This may be something that many people are doing everyday but I
>> did it only once some years ago. I am running S-35 labelled
>> immunoprecipitates by SDS-PAGE. I cannot detect anything by
>> drying the gel and doing autoradiography. So I will try Fluorography
>> which enhances detection on X-Ray films. Can somebody send me
>> his/her protocol for processing the gel after running it.
>> Maniatis talks about PPO and salisylate methods but does not
>> describe the details ( the actual step by step protocol ).
>> Thanks for taking the time and replying.
>>
>> Zafer
>
>Get in touch with Amersham.  They do a product call 'Amplify' which is
>very quick and easy.  You basically fix your gel (30 mins); soak in
>'Amplify' (15-30 minutes); dry and expose the gel.
>Amersham have also written a few reviews which you get from them.  Review
>23 is called "radioisotpoe detection by fluorography and intensfying
>screens".  It is very good and is worth getting hold of for the procedures
>within.
>
>In short if I wre you I would use Amplify.


__
Jonathan B. Marder                 '
Department of Agricultural Botany  |     Internet: MARDER at agri.huji.ac.il
The Hebrew University of Jerusalem | /\/
Faculty of Agriculture             |/  \ Phone:    (08 or +9728) 481918
P.O.Box 12, Rehovot 76100, ISRAEL  /     Fax:      (08 or +9728) 467763



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