Chromatography Forum

Hi,

I am planning to separate/enrich phosphorylated peptides (proteins) by the means of HPLC. Anyone to recommend a suitable column containing a phase such as an IMAC-phase or any other appropriate material?

Any other comment/hint in respect to the above topic is very much appreciated.

Thanx alot.

SMC,

IMAC in the ferric form is a very good way to classify phosphopeptides, but is more hit-and-miss with phosphoproteins. I don't know of any IMAC product in an HPLC format. (We are beta testing one, but it is not yet a "product." Contact me privately if you are interested.)

Sigma and BioRad have ferric-IMAC resins you can buy for use with extraction tubes. I've tried the Sigma product with tryptic peptides and it worked.

You didn't say how you plan to further characterize the peptides. There is a nice paper by Annan, et.al., Analyt. Chem. 2001, 73, 393-404.

HI SMC,

I've got information and application data on our Protecol-IMAC trap column and it's use to separate/enrich phosphorylated peptides . Please contact me if you're interested.

Cheers

Hugh
SGE Europe Ltd