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Protein Expression & Purification


Products >  Protein_Expression_and_Purification >  His-Tagged_Protein_Purification >  Large_Volume

His-Tagged Protein Purification from Large Lysate Volumes—Capturem His-Tagged Purification Large Volume

Capturem His-Tagged Purification Large Volume provides a revolutionary solution for rapidly purifying and concentrating proteins from large volumes of mammalian or bacterial cell lysate under a wide range of conditions—by taking advantage of a recently developed technology using modified Ni-NTA membranes. It consists of single-use, disposable bottle-top membrane units for simple, rapid, vacuum-based purification of his-tagged proteins at room temperature from up to 500 ml (at a time) of clarified lysate.

Isolate his-tagged proteins of interest with a purification system that is:
  • Fast and convenient: works in 10–30 min at room temperature instead of 6–10 hr in the cold room
  • High yield: up to 25 mg/unit, using a novel membrane technology instead of resin
  • Highly concentrated eluate: up to 2.0 mg/ml
  • Flexible: purify from mammalian or bacterial cells
Where resin columns fall short, this new system provides high capacity and purity in just 10–30 min, from loading the sample through elution. Novel nylon membranes have been assembled into bottle-top units, creating a no-waiting, room-temperature workflow compatible with both mammalian and bacterial cells. Exceptional performance is consistent under a wide range of sample preparations, including native and denaturing conditions, the presence of additives (e.g., βME, EDTA, DTT, glycerol, TCEP, etc.; see table below), and tandem purification of his-tagged proteins.

The modified nylon membranes have a small bed volume, yet still produce high-purity, concentrated protein due to the chemically enhanced surface area of the pores. With this technology, protein binding capacity is comparable to, or better than, resins that bind 75 mg or more per cm3 of membrane.

Each bottle-top unit has a sample capacity of up to 500 ml. (For proteins expressed at low levels, an additional 500 ml of cleared cell culture supernatant may be loaded into the unit.) Each unit produces up to 25 mg of protein at a concentration of ~ 1.0 mg/ml (depending on protein size and expression level) using the recommended amount of elution buffer, and up to 2.0 mg/ml if a smaller amount of elution buffer is used. The short residence time greatly reduces the possibility of protein degradation, aggregation, or loss of activity, giving downstream applications a higher probability of success.

Since the Capturem His-Tagged Purification Large Volume bottle-top units are designed for 33-mm diameter thread bottles, a Bottle Thread Adaptor is included to ensure compatibility with 45-mm diameter thread bottles.

  At-A-Glance   Documents   Images & Data

Features

  • No-waiting workflow: 10- to 30-min, room-temperature protocol
  • High purity and yield: up to 25 mg/unit (depending on protein size and expression level), at a concentration of up to 2.0 mg/ml
  • Wide range of conditions: native and denaturing conditions; with common additives (e.g., EDTA, DTT, βME, glycerol, TCEP, etc.)
  • Lysis buffer compatibility: xTractor Buffer, buffers from other vendors
  • Different cell systems: easy purification from mammalian and bacterial cells

Applications

  • Biochemical/enzymatic assays
  • Pull-down or other protein interaction assays
  • Confirmation screening
  • Protein/ligand affinity analysis
  • Crystallography or other structural analyses
  • Antibody production
  • Toxicology studies
  • Protein transfer to live cells

Reagents Compatible with Capturem His-Tagged Purification Large Volume

Reagent Compatibility*
MOPS200 mM
HEPES200 mM
Tris200 mM
EDTA10 mM
Beta-mercaptoethanol30 mM
DTT10 mM
TCEP5 mM
Guanidine-HCl6 M
Urea8 M
Nonionic detergent (Triton X-100)2%
Nonionic detergent (Tween 20)2%
Anionic detergent (SDS)1%
Arginine500 mM
Glycine100 mM
Histidine20 mM
Sodium chloride2 M
Imidazole40 mM
Glycerol10%

* Compatible with concentrations up to the amounts shown. These are the maximum amounts that have been tested.


 
 
Products
New Offers Cat. # Product Contents Size Price License Units Select
635724 Capturem™ His-Tagged Purification Large Volume 4 Units (2 Units per box) $396.00 License Statements
 

 

 
 

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