Dear Amit, You are working with a membrane protein and the use of SDS (harsh detergent) often makes these proteins to oligomerise. Boiling your sample is not advisable, it might make it worse.
I’m not sure why you would like to “see” a monomer in the gel but if you really would like to know what is the oligomeric state of you protein in solution, I would advise you to do a SEC-MALS (size exclusion chromatography - multi angle light scattering) run of your sample. Best, Isabel --------------------------------------------------------- Dr Isabel Moraes Membrane Protein Laboratory Group Leader Diamond Light Source Ltd, Harwell Science and Innovation Campus, Oxfordshire, OX11 ODE, UK --------------------------------------------------------- On 21 Feb 2017, at 22:22, amit gaur <cdriamitg...@gmail.com<mailto:cdriamitg...@gmail.com>> wrote: Hi all, I am trying to purify a potassium ion channel from insect cell using baculovirus expression system. I am not seeing monomer of this protein in SDS instead a dimer appears.So,I increased DTT in SDS buffer but no change and dimer was intact. In size exclusion this protein appeared as a tetramer which is common oligomerizaton of potassium channel family with GYG motif. Can any body suggest what should I do in this case? Thanks and regards, -- Dr. Amit Gaur Post Doctoral Researcher PI: Dr. Ji-Fang Zhang Thomas Jefferson University 1020, Locust Street, Suite 418 Philadelphia, PA 19107 -- This e-mail and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorised recipient of the addressee please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to the e-mail. Any opinions expressed within this e-mail are those of the individual and not necessarily of Diamond Light Source Ltd. Diamond Light Source Ltd. cannot guarantee that this e-mail or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. Diamond Light Source Limited (company no. 4375679). Registered in England and Wales with its registered office at Diamond House, Harwell Science and Innovation Campus, Didcot, Oxfordshire, OX11 0DE, United Kingdom
