Removal of SDS

Ashok M S lambu at biochem.iisc.ernet.in
Tue Dec 12 10:54:41 EST 2000


 Hi,  here is a protocol to remove SDs from electro eluted protein.
 After elution lyophilize the sample and resupend in 100ul of water.
2. Add 400ul of methanol and vortex it, and spin at 9000g/10 sec.
3. add 100ul of chloroform (equilbrated with water), vortex and spin for
   10 sec at 9000g.
4.Add 300ul of water to effect phase seperation, vortex and spin 9000g
  for 1 min.
5. precipitated protein will be in the interface.
6. remove the upper phase andadd 300 ul of methanol vortex and spin at
9000g for 2 min
7. Remove the sup and wash with 95% methnaol, spin 9000g 2 min and air
drythe pellet.

 i hope this helps and i am sorry for replying to your query late.
 i have a protocol for estimating SDS in protein samplefor  which i have
 to hunt for in my books. if u need it let me know

Cheers
Ashok.



On 17 Nov 2000, Frankie wrote:

> We purified an antibody in SDS-PAGE gel and recovered the antibody from
> the gel using electro-elution.  We found that the recovery rate was poor
> unless SDS containing elution buffer was used.  However, the presence of
> SDS in the elutant is not desirable in the following experiment.  Can
> anyone suggest us an effective way to remove the SDS from the elutant or
> any effective non-SDS elution buffer for electro-elution.  Thank you.
> 
>  TAM Frankie
> 
> 
> ---
> 
> 

-- 
    "I may not be totally perfect, but parts of me are excellent."
                                                  
                                                     Ashleigh Brilliant
                                                    
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Ashok M. S                              Phone   
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Department of Biochemistry            Hostel : 309-2539 (R-67) 
Indian Institute of Science            email : lambu at biochem.iisc.ernet.in 
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