5 replies to this topic

[avalon]

Hi all,  

Can anyone recommend a non-reducing protein ladder?

[mdfenko]

ge healthcare sells a couple of sets of markers that may be what you want (did you mean non-denatured?).

ge healthcare protein standards

protein and peptide mw markers

(the lmw-sds markers do not contain sds or dtt or 2-me, they can be used for native page)

Edited by mdfenko, 16 May 2011 - 10:52 AM.

[avalon]

mdfenko, on 16 May 2011 - 10:50 AM, said:

ge healthcare sells a couple of sets of markers that may be what you want (did you mean non-denatured?).

ge healthcare protein standards

protein and peptide mw markers

(the lmw-sds markers do not contain sds or dtt or 2-me, they can be used for native page)

Thanks for replying mdfenko,  I actually wont be running native gels ( I will be doing sds page), but am trying to detect a biotinylated protein after IP with a specific antibody.....just trying to confirm specificity of my IP without using radioactivity.  I havent had much success so far and someone pointed out that the reducing agents in my ladder were probably cleaving the biotin off the protein (I should have used a different biotin to start with).  So I guess I just need a ladder without DTT or BME really but a lot of the companies dont seem to specify whether these are present or not.

[mdfenko]

the lmw standard does not contain reducing agent and is good for sds-page.

[avalon]

mdfenko, on 17 May 2011 - 07:16 AM, said:

the lmw standard does not contain reducing agent and is good for sds-page.

Thank you!

[Jennifer415]

Why do you care if your ladder is reduced or not............it runs separately from the other protein lanes!  doubtful that the small amount of reducing agent is cleaving biotin in your protein lanes.