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NEBExpress Ni Spin Columns

NEBExpress Ni Spin Columnsカタログ番号:S1427

カタログ番号

サイズ

濃度

価格(税別)

保存温度

S1427S 10 columns 0.1 ml bed volume ¥18,200 4C
S1427L 25 columns 0.1 ml bed volume ¥36,400 4C

製品カテゴリ>グループ

  • タンパク質発現&精製>PURExpress (無細胞タンパク質合成システム)

備考

  • 備考:価格改定 (2023年4月1日)

特徴

・すぐに使えるニッケル・スピンカラム

・精製に必要なバッファーが付属

・わずか 15 分間で ≥1 mg の His タグタンパク質を精製

・>95 % の高純度精製

・EDTA や界面活性剤が入ったライセートも使用可能

・ネイティブ、変性条件の両方で精製可能

 

説明:

小スケールの His タグタンパク質の精製に便利なニッケル・スピンカラムである。わずか 15 分間で高純度で精製ができる。

 

サポートマトリックス:

Spherical, agarose based microparticles ranging in size from 10-100 μm.


Figure 1: NEBExpress Ni Spin Column Quick Start Protocol

image

 

 

Figure 2: Purification of His-tagged fusion proteins using NEBExpress Ni Spin Columns

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NEBExpress Ni Spin Columns were equilibrated and loaded with 500 μl of crude extract from E. coli containing a plasmid that expresses either His6-GluRS or His6-NDK fusion protein. Columns were washed, eluted and analyzed by SDS-PAGE to confirm low non-specific binding of extract proteins and high iso­lation of target. Marker (M) contains Color Prestained Protein Standard (NEB #P7719), Lane 1: GluRS protein load, Lane 2: GluRS flow through, Lane 3: GluRS 1st elution, Lane 4: GluRS 2nd elution, Lane 5: NDK protein Load, Lane 6: NDK flow through, Lane 7: NDK 1st elution, Lane 8: NDK 2nd elution.


付属試薬:

・2X IMAC Buffer

・2M Imidazole

製品情報

保存温度:

4°C


保存条件:

20% Ethanol


結合能:

≥ 1 mg His タグタンパク質(カラムあたり、タンパク質の種類によって異なる)

メモ

1.Do not freeze.


2.Several E. coli proteins are capable of binding immobilized metal affinity matrices with moderate to high affinity. The most common contaminants include SlyD (28kDa), GlmS (67kDa), ArnA (74kDa) and carbonic anhydrase (25 kDa) (1). The named contaminants may be eliminated from the Ni column elution fraction by employing NiCo21(DE3) (NEB# C2529H) as the expression host and then performing a second binding step with chitin magnetic beads (NEB# E8036) (2). 


3.Protein yield and purity are dependent upon the expression level, conformation and solubility characteristics of the recombinant fusion protein.  For best results, estimate the expression level of the His-tagged protein of interest by first running a sample of the crude lysate on an SDS-PAGE gel. 


4.Unlike most IMAC resins, the unique chemistry employed in NEBExpress™ Ni Resin provides chemical tolerance to chelating agents such as EDTA and reducing agents such as DTT.  Please consult the chemical compatibility table for more information. 


5.Polyhistidine tags do not typically compromise the biological protein function and are not considered immunogenic, however if cleavage of the His-tag is necessary we recommend using TEV Protease (NEB# P8112).


6.Ni resin yields highly variable binding capacities dependent on the target and conditions. The binding capacity value (≥ 1 mg/column) for this product was determined by performing a mock purification from crude cell lysates expressing a His-tagged target protein.