Product selection Protino® Ni-TED / IDA

 

Structure of TED and IDA in complex with Ni2+

	Protino® Ni-TED	Protino® Ni-IDA
Matrix	Macroporous silica
Chelating group	TED	IDA
Binding sites Ni2+ to His Tag	1	3
Binding sites ligand to Ni2+	5	3
Ligand density	high	low
Binding capacity	10 mg/g resin	20 mg/g resin
Specification	High binding specificity
	Less unspecific binding of contaminating proteins compared to Ni-NTA and Ni-IDA Agarose
	Elution at low imidazole concentrations possible
	High stability against reducing / chelating agents	High protein yield / recovery even from diluted samples
	Low metal leaching	High protein concentration
	High protein purity
Available formats	Protino® Ni-TED Packed Columns	Protino® Ni-IDA Packed Columns
	Protino® Ni-TED Resin	Protino® Ni-IDA Resin
		Protino® 96 Ni-IDA

Reagent compatibility Protino® Ni-TED / IDA

Reagent	Effect		Comments
Sodium phosphate	Used in LEW and Elution buffer in order to buffer the solutions at pH 8		50TED/IDA mM is recommended. The pH of any buffer should be adjusted to 8, although in some cases a pH between 7 and 8 can be used
Tris	Coordinates with Ni2+ ions, causing a decrease in capacity		10TED/IDA  mM may be used, sodium phosphate buffer is recommended
Sodium Chloride	Prevents ionic interactions and therefore unspecific binding		Up to 2TED/IDA  M can be used, at least 0.3 M should be used
Imidazole	Binds to immobilized Ni2+ ions and competes with the polyhistidine-tagged proteins		Should not be included in LEW buffer
Urea	Solubilizes protein		Use 8TED/IDA  M for purification under denaturing conditions
GuHCl	Solubilizes protein		Up to 6TED/IDA  M can be used
ß-mercaptoethanol	Prevents formation of disulfide bonds; Can reduce Ni2+ ions at higher concentrations		Up to 50TED/IDA  mM in samples has been used successfully in some cases
DTT, DTE	Can reduce Ni2+ ions at higher concentrations		Up to 10TED/IDA  mM in samples has been used successfully in some cases
Glutathione reduced	Can reduce Ni2+ ions at higher concentrations		Up to 30TED/IDA  mM in samples has been used successfully in some cases
Glycerol	Prevents hydrophobic interactions between proteins		Up to 50TED/IDA  % can be used.
EDTA	Coordinates with Ni2+ ions, causing a decrease in capacity at higher concentrations		Up to 10TED/1IDA mM in samples has been used successfully in some cases
Ethanol	Prevents hydrophobic interactions between proteins		Up to 20TED/IDA % can be used; Ethanol may precipitate proteins, causing low flow rates and column clogging
SDS	Interacts with Ni2+ ions, causing a decrease in capacity		Not recommended, but up to 0.2TED/0.5IDA % has been used successfully in some cases
Triton, Tween	Removes background proteins		Up to 2TED/IDA  % can be used
Do not use buffers with pH > 8.4 since silica dissolves in solutions of high pH.