Editing HTNCre HIC Purification (section)

== Method 2(high salt condition):  ==

1.Equlibrate column with buffer 2M (NH4)2SO4 +20mM Tris-HCl 

2.Load sample 0.5ml buffer (2M (NH4)2SO4 +20mM Tris-HCl) +0.5ml sample 

3.Wash colume with 5 ml buffer 2M (NH4)2SO4 +20mM Tris-HCl into 5 tubes 

4.Washcolume with 5 ml buffer 2M NaCl/20mM Tris-HCl /10%glycerol into 5 tubes<br> 

5.Wash colume with 5 ml buffer 1M NaCl/20mM Tris-HCl /10%glycerol into 5 tubes<br> 

4.Elute with 5 ml 1*PBS/10%glycerol into 5 tubes 

5.Test protein with Brandford to observe which tube protein is in 

result: protein all exist in last 10 wash wells and 5 elute wells, but very little in each well,protein lose too much ,this method can not be used for protein purification <br> 

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[[Category:Protein_Purification]]