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I am new to HPLC... Pl. help me!!!

Posted: Tue Apr 08, 2008 10:06 pm
by sechau1
:(
Hi, I am a newbie to HPLC. I have to use SEC-HPLC for analytical purpose to see %monomer, %aggregate etc from a purified Ab (M.Wt: 150KDa). We have an old Waters 2695 in lab. Could you please suggest what column (if possible brand/model) that I should use? Also, do I need several columns?
Thanks in advance.
A lost Grad student
:cry:

Posted: Wed Apr 09, 2008 12:49 am
by Uwe Neue
Either use a Protein-Pak 300 7.5 x 300 or a BioSuite SEC 250 7.8 x 300 column (both Waters) at a flow rate of 1 mL/min, phosphate buffer pH 7, about 25 mM plus 100 mM NaCl. You can separate the monomer from the dimer on a single column.