How to get rid of DNA in cell lysate

Pow Joshi pow.joshi at gmail.com
Mon Jun 26 21:07:51 EST 2006


> I lysed the fibroblast with 1% triton in PBS by freeze thawing. After
> loading on the SDS gel I don't see any protein in the fraction. My antibody
> is specific and it works well with the cell lysate containing PBS alone.

.... were you able to see any protein bands at all with PonceauS after
the transfer, in the fibroblast lane? .
you could also add a protease inhibitor cocktail containing PMSF,
leupeptin, pepstatin and aprotinin, if you are'nt already doing so....
>
>   I was wondering if the Presence of DNA in cell lysate mask the appearance
> of the protein on SDS gel and how to get rid of that.

you could also pass the lysate through a syringe 23 and 26 guage
needle about 10 times each to shear the DNA and then spin down the
samples, and use the suprenatant for SDS PAGE.
>

Or the problem is something else.
>   Any help will be appreciated.


>


On 6/25/06, Maryam <mzargh at yahoo.ca> wrote:
> I lysed the fibroblast with 1% triton in PBS by freeze thawing. After
> loading on the SDS gel I don't see any protein in the fraction. My antibody
> is specific and it works well with the cell lysate containing PBS alone.
>
>   I was wondering if the Presence of DNA in cell lysate mask the appearance
> of the protein on SDS gel and how to get rid of that.
>   Or the problem is something else.
>   Any help will be appreciated.
>
>  		
> ---------------------------------
> Be smarter than spam. See how smart SpamGuard is at giving junk email the
> boot with the All-new Yahoo! Mail
> _______________________________________________
> Methods mailing list
> Methods at net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>



More information about the Methods mailing list