ProteinIso Ni NTA Resin His Tag Protein

Product Overview

Description

ProteinIso Ni-NTA Resin

ProteinIso^TM Ni-NTA Resin is used for one-step purification of Histagged proteins. The His-tagged proteins bind to Ni²⁺ cations, which are immobilized on the Ni-NTA resin by 4 metal-chelating sites. After unbound proteins are washed away, the target proteins are recovered by gradient elution. It is suitable for both native and denatured protein purification.

 

Cat. No.
Specification	DP101-01	5 ml
	DP101-02	25 ml
Storage:	at 2~8oC (20% ethanol) for two years
Specification	Samples should be centrifuged and filtrated with 0.45 μm filter before loading.
	Equilibration Buffer for soluble protein: 300 mM NaCl, 50 mM sodium phosphate buffer, 10 mM imidazole, 10 mM Tris-Cl pH 8.0
	Equilibration Buffer for inclusion body: 6 M GuHCl, 100 mM sodium phosphate buffer; or 8 M urea, 100 mM sodium phosphate buffer, 10 mM Tris-Cl pH 8.0

 

Resin Specifications

References	Specifications
Matrix	6% cross-linked agarose
Ligand	NTA
Shape	sphere
Pore size	45-165 μm
Binding capacity	10-20 mg proteins/ml wet gel
Recommended flow rate	<300 cm/h
Highest resistance of atmospheric pressure	0.3 Mpa
pH stability	3-13

 

Procedures

 

1. Prepare Ni-NTA purification column

(1) Resuspend the Ni-NTA resin in its bottle by inverting.

(2) Transfer the resin into a purification column. Allow the resin to settle.

(3) Equilibrate the column with 5~10 bed volume of equilibration buffer.

2. Prepare samples

To avoid blocking column, samples should be centrifuged and filtrated before loading.

3. Load samples and wash

Load samples and wash with 5~10 bed volume of equilibration buffer and collect the flow-through in one tube.

4. Elute

Elute target protein with different concentration imidazole or different pH equilibration buffer.

5. Resin Regeneration

(1) 2 bed volume of 6 M GuHCl, 0.2 M acetic acid

(2) 5 bed volume of deionized water

(3) 3 bed volume of 2% SDS

(4) 1 bed volume of 25% ethanol

(5) 1 bed volume of 50% ethanol

(6) 1 bed volume of 75% ethanol

(7) 5 bed volume of 100% ethanol

(8) 1 bed volume of 75% ethanol

(9) 1 bed volume of 50% ethanol

(10) 1 bed volume of 25% ethanol

(11) 1 bed volume of deionized water

(12) 5 bed volume of 100 mM EDTA, pH 8.0

(13) 10 bed volume of deionized water

(14) 5 bed volume of 100 mM NiSO₄

 

 

 

 

 

 

 

 

 

 

CITATIONS

Cao H, 2013. FCHSD1 and FCHSD2 Are Expressed in Hair Cell Stereocilia and Cuticular Plate and Regulate Actin Polymerization In Vitro. 8(2):e56516. PLoS One. IF=3.73. PMID: 23437151.

 

Wang L, et al. 2011. Cloning of exoinulinase gene from Penicillium janthinellum strain B01 and its high-level expression in Pichia pastoris. J Appl Microbiol. IF=2.196. PMID: 21895898.

 

Huang H, 2013. Cloning, expression and characterization of a phosphoglucomutase/phosphomannomutase from sphingan-producing Sphingomonas sanxanigenens. 35(8):1265-70. Biotechnol Lett. IF=1.853. PMID: 23546942.