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Ni-NTA Fast Start Kit

For purification and detection of recombinant His-tagged proteins from E. coli lysates

  • Ideal for researchers new to protein science
  • Everything required for efficient purification in one kit
  • Easy-to-follow protocols and straightforward processing
  • Up to 25 mg protein per column in as little as 90 minutes
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)
$381.00
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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.
0
Highly pure His-tagged protein.
1
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.
Performance
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.
Principle

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.

Procedure
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.
Applications

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
Features
Specifications
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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