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 Multi-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