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Ni-NTA Fast Start Kit

 E. coli lysates에서 recombinant 6xHis-tagged proteins를 분리 정제 및 검출할 수 있습니다
  • 단백질 실험 초보자에게 적합
  • 하나의 kit으로 효율적인 분리 정제에 필요한 모든것
  • 따르기 쉬운 프로토콜과 간단한 처리과정
  • 90분 안에 column당 10 mg의 protein 분리 정제 가능
The Ni-NTA Fast Start Kit provides everything needed for fast, efficient purification of His-tagged proteins from cleared E.coli lysates, including prefilled Ni-NTA columns. Buffers supplied in the kit enable proteins to be purified either under native or denaturing conditions. The kit also contains an Anti-His antibody for detection of expressed His-tagged proteins.
Cat No./ID: 30600
Ni-NTA Fast Start Kit (6)
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For purification and detection of six 6xHis-tagged protein preps: 6 x Fast Start Columns, Penta·His Antibody, Buffers and Reagents
The Ni-NTA Fast Start Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Highly pure His-tagged protein.
High purity proteins under native or denaturing conditions.
Coomassie-stained SDS-PAGE gels showing  [A]  6xHis-tagged thioredoxin purified under denaturing conditions, and  [B]  6xHis-tagged green fluorescent protein (GFP) purified under native conditions. M: markers; CL: cell lysate; FT: flow-through fraction; Wash: wash fractions; E1 & E2: elution fractions. 5 µl of each fraction was loaded on to the gel.
The Ni-NTA Fast Start Kit provides everything required for fast, efficient purification of His-tagged proteins from cleared E. coli lysates. Buffers supplied in the kit enable proteins to be purified either under native or denaturing conditions (see figure High purity proteins under native or denaturing conditions) Simple, straightforward protocols and ready-to-use reagents make the Ni-NTA Fast Start Kit an ideal starting point for scientists looking to expand their research into the field of protein expression, but who are unfamiliar with protein purification procedures.

The Ni-NTA Fast Start Kit is based on the remarkable selectivity of patented Ni-NTA (nickel-nitrilotriacetic acid) resin for proteins containing an affinity tag of six or more histidine residues (consecutive or alternating) — the His tag. This technology allows one-step purification of almost any His-tagged protein under native or denaturing conditions. NTA, which has four chelation sites for nickel ions, binds nickel more tightly than metal-chelating purification systems that only have three sites available for interaction with metal ions. The extra chelation site prevents nickel-ion leaching and results in protein preparations with higher purity than those obtained using other metal-chelating purification systems. The His tag also forms the epitope recognized by the Penta·His Antibody supplied with the kit.

Cells are lysed in the supplied lysis buffer and centrifuged to obtain a cleared lysate. This lysate is applied to a Fast Start Column, where His-tagged proteins bind with high affinity and untagged proteins pass through. After washing, purified proteins are eluted using buffers containing imidazole (native conditions) or of low pH (denaturing conditions) (see figure Highly pure 6xHis-tagged protein). The purification procedure and protein yield can be followed by SDS-PAGE, western blotting and immunodetection using the supplied Penta·His Antibody.

The Ni-NTA Fast Start Kit provides reliable, one-step purification of 6xHis-tagged proteins suitable for any application, including:

Functional studies
Immunization to produce antibodies
Assays involving protein–protein and protein–DNA interactions
Structural investigations
Applications Proteomics
Binding capacity 5–20 mg/ml
Gravity flow or spin column Gravity flow
Processing Manual
Special feature Ni-NTA technology
Start material Cell lysate
Support/matrix Ni-NTA matrix
Tag 6xHis tag
Yield <10 mg protein per column

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