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
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
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."
Ashok M. S Phone
Research Scholar Lab : 309-2540, 360-1492
Department of Biochemistry Hostel : 309-2539 (R-67)
Indian Institute of Science email : lambu at biochem.iisc.ernet.in
Bangalore 560 012. INDIA
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