GE Life Science Products

Protein Purification

  Affinity Tagging




Affinity tagging has revolutionized protein purification by allowing generic purification approaches, and today many proteins can be purified very easily and efficiently.

In research laboratories, protein purification is performed in scales from micrograms and milligrams, while bioprocessing at industry scale can produce kilograms or even tons. Preparative purification of proteins commonly involves chromatography, GE offers a wide range of chromatography media to purify proteins using either manual or automated methods. Prepacked formats include gravity columns, spin columns, 96-well plates, as well as prepacked columns for use with automated chromatography systems.

For step-by-step protocols and practical tips about protein purification, download one of the in-depth handbooks (see Handbooks tab below).

 Histidine-Tagged Protein Purification

Histidine tags are widely used because they are small and rarely interfere with the function, activity, or structure of target proteins. Immobilized metal ion affinity chromatography (IMAC) is the most common method for purifying histidine-tagged proteins. IMAC chromatography media charged with divalent metal ions such as nickel or cobalt selectively retain histidine-tagged proteins and allow for the purification of insoluble histidine-tagged proteins from inclusion bodies when denaturing conditions are used. Successful IMAC purification gives a high yield of pure and active target protein.

Since many proteins have intrinsic histidine and/or cysteine amino acid residues, other nonspecific proteins can bind to the IMAC media together with the target protein. In such cases, it is often necessary to optimize binding, wash, and elution conditions by varying the concentration of imidazole in these solutions. Increasing the concentration of imidazole in the binding and wash buffers generally decreases nonspecific binding, whereas lower concentrations give stronger affinity interaction. The key is finding the right balance.

Sigma-Aldrich offers GE products for fast and convenient histidine-tagged recombinant protein purification using immobilized metal ion affinity chromatography (IMAC). Ni Sepharose media are precharged with nickel ions (Ni2+) and TALON® Superflow™ medium is precharged with cobalt ions (Co2+). Ni Sepharose excel is especially suitable for purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants.

IMAC Sepharose media are uncharged and can be used with a metal ion of your choice. Our range of IMAC media is available in a variety of formats including columns, 96-well plates, and magnetic beads.

A polyhistidine tag is an amino acid motif in proteins that consists of at least five histidine (His) residues linked to the N- or C-terminus of the protein. The (histidine)6-tag, using six His residues, is popular based on its small size, strong metal ion binding, and ability to bind under denaturing conditions.

IMAC media charged with Ni2+ and Co2+ are the most commonly used methods for the purification of histidine-tagged proteins. However, in some cases, other metal ions may be more suitable, for example copper ( Cu2+) or zinc (Zn2+). In these cases, uncharged IMAC media can be conveniently charged with the metal ion of your choice.

 Cobalt-based TALON® Protein Purification

Cobalt-based TALON® products are the first choice when the purity of your histidine-tagged protein is more important than yield. TALON® Superflow™ binds polyhistidine-tagged proteins with an enhanced specificity than nickel-based media due to the structure of the cobalt ligand. The result is that TALON® binds fewer unwanted proteins.

Formats for purification without a chromatography system include:

  • His GraviTrap TALON® columns
  • His SpinTrap TALON® columns
  • His MultiTrap TALON® 96-well filter plates (can be used with a centrifuge or vacuum)
  His MultiTrap TALON® is a ready-to-use 96-well filter plate for high reproducibility in expression screening and small-scale parallel purification of histidine-tagged proteins.
  His SpinTrap TALON® is a single-use column for rapid screening with small samples of histidine-tagged proteins.
  His GraviTrap TALON® is a prepacked gravity flow column for convenient manual purification of histidine-tagged proteins.
  TALON® Superflow™ is a cobalt-based IMAC medium offering different selectivity compared to nickel-charged media.
 
Prepacked columns suitable for automated chromatography systems:
HiTrap TALON® Crude
1ml and 5ml columns allow for rapid separations and easy scale-up with a minimum of sample preparation and equipment.
     • GE29048565
     • GE28953766
     • GE28953805
     • GE28953767
     • GE28953809

All TALON® products are suitable for purification under native or denaturing conditions. They typically require less imidazole than nickel-based media for washing and elution, which preserves the structure and function of sensitive histidine-tagged proteins.

 His Buffers

  His Buffer Kit


 His Detection

  Anti-His Antibody

 


 Sepharose

Ni Sepharose High Performance (HP), Ni Sepharose 6 Fast Flow (FF), and Ni Sepharose excel chromatography media are precharged with Ni2+ ions for convenience. GE’s immobilized metal ion chromatography (IMAC) media provide high binding capacity or high yield depending on your needs.

Disposable Columns Prepacked with Sepharose

Single use columns prepacked with precharged Ni Sepharose media.

  His GraviTrap™ is a prepacked, single-use column for purification of histidine-tagged proteins by immobilized metal affinity chromatography (IMAC). Column gives fast and simple gravity-flow purifications without any need for a purification.

 

  His GraviTrap Kit

 

  His SpinTrap™ is a prepacked, single-use spin column for purifying histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC)

 

  His Spintrap Kit is a prepacked, single-use spin column for purifying histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC).

 

HiScreen Columns Prepacked with Sepharose

HiScreen is a 4.7ml ready-to-use column, prepacked with precharged Ni Sepahrose media.

  HiScreen FF – HiScreen columns prepacked with Ni Sepharose 6 Fast Flow for method optimization and parameter screening for purification of histidine-tagged proteins with immobilized metal ion affinity chromatography (IMAC).

 

MultiTrap Filter Plates Prepacked with Sepharose

MultiTrap is a ready-to-use 96-well filter plate for high reproducibility.

    His MultiTrap Fast Flow (FF) and His MultiTrap High Performance – His MultiTrap FF and His MultiTrap HP are prepacked disposable 96-well plates for reproducible high throughput parallel purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).

 

  His MultiTrap Fast Flow (FF)

 

  His MultiTrap High Performance

 

HiPrep Columns Prepacked with Sepharose

HiPrep is a 20ml ready-to-use column prepacked with precharged Ni Sepahrose media.

  HiPrep FF 16/10 is a ready-to-use column, prepacked with precharged Ni Sepharose 6 Fast Flow. the column is an excellent choice for preparative, scale-up purification of histidine-tagged recombinant proteins.

 

HiTrap Columns Prepacked with Sepharose

 

  HisTrap FF is a ready-to-use column, prepacked with precharged Ni Sepharose 6 Fast Flow for preparative purification of histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).
     • GE17531901
     • GE17531902
     • GE17525501
     • GE17525502

 

  HisTrap Excel columns are prepacked with Ni Sepharose excel affinity media for capture and purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants by immobilized metal ion affinity chromatography (IMAC).
     • GE29048586
     • GE17371205
     • GE17371206

 

  HisTrap FF Crude columns offer the convenience and simple, one-step purification of histidine-tagged proteins directly from homogenized, unclarified cell lysate by immobilized metal ion affinity chromatography (IMAC).
     • GE29048631
     • GE11000458
     • GE11000459
     • GE17528601
     • GE17528602
     • GE28401477

 

  HisTrap HP columns are prepacked with Ni Sepharose High Performance and designed for simple, high-resolution purification of histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC).
     • GE29051021
     • GE17524701
     • GE17524705
     • GE17524801
     • GE17524802
     • GE17524805

 

Sepharose Chromatography Media

Ni Sepharose media conveniently precharged with Ni2+ ions.

  Ni Sepharose Excel affinity media for capture and purification of histidine-tagged proteins secreted into eukaryotic cell culture supernatants by immobilized metal ion affinity chromatography (IMAC).
     • GE17371202
     • GE17371203

 

  His Mag Sepharose Ni is magnetic beads designed to simplify small-scale purification and expression screening of histidine-tagged proteins.
     • GE28967388
     • GE28967390
     • GE28979917

 

  Ni Sepharose High Perfomance affinity media for high-resolution, high capacity purification of histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC).
     • GE17526801
     • GE17526802

 

  Ni Sepharose 6 Fast Flow is a BioProcess medium that combines the advantages of Ni2 - for purification of histidine-tagged proteins, with the well industrially established properties of the Past Flow platform.
     • GE17531806
     • GE17531801
     • GE17531802
     • GE17531803

 

  His Mag Sepahrose Excel magnetics beads are designed for small-scale purification and screening of histidine -tagged proteins secreted in eukaryotic cell culture.
     • GE17371220
     • GE17371221
     • GE17371222

 

 GST-Tagged Protein Purification

Glutathione Sepharose chromatography media (resins) are available in lab packs, prepacked GSTrap and GSPrep columns, and 96-well plates. We also offer GST Purification Modules for purification of bacterial lysates. GST SpinTrap module includes 50 prepacked Glutathione Sepharose 4B SpinTrap columns as well as the buffers needed for purification. RediPack GST Purification Module includes gravity flow columns and buffer for purifying up to 20 mg protein.

Glutathione S-transferase (GST) Gene Fusion System is a versatile system for the expression, purification, and detection of GST-tagged proteins produced in E. coli. Purification of GST-tagged proteins can be performed under very mild conditions, which preserves the function and antigenicity of the target protein.

The GST tag may also increase the solubility and stability of the target protein. The GST tag is large with a relative molecular mass (Mr) of 26000. A specific cleavage sequence allows simple removal of the GST tag with the use of different proteases after purification.

Typical features of GST-tagged proteins include high binding specificity to glutathione ligands on Glutathione Sepharose chromatography media, resulting in greater than 90% purity in one step of the eluted target molecule.

GST is an enzyme found in most organisms. The functional protein is a dimer. The tag consists of 220 amino acid residues, making the tagged protein more soluble as well as simplifying its purification and detection. However, removal of GST tag from the target protein is often necessary due to its large size. This can be performed with PreScission Protease which is optimized for low-temperature cleavage.

The purification method is based on the high affinity of GST for glutathione. When applied to the affinity medium, GST-tagged proteins bind to the glutathione ligand, and impurities are removed by washing with binding buffer. Tagged proteins are then eluted from the chromatography medium under mild, non-denaturing conditions that preserve both protein structure and function. GST Buffer Kit contains prepared buffer concentrates for binding, washing, and elution of GST-tagged proteins.

 GST Sepharose

Glutathione Sepharose High Performance (HP), 4 Fast Flow (FF) and 4B are media for purification of Glutathione S-Transferase (GST) tagged proteins. These media offer different characteristics depending on your needs.

Disposable Columns Prepacked with Sepharose

Single-use columns prepacked with Glutathione Sepharose media.

  GST Bulk Kit is designed for small-scale manual purification of GST-tagged proteins using either column chromatography or batch method. Facilitates in optimization.

 

  GST GraviTrap™ contains 10 disposable columns prepacked with 2 ml of Glutathione Sepharose 4B, sufficient for purification of up to 20 mg of fusion protein.

 

  GST SpinTrap™ columns are designed for rapid small-scale purification of GST-tagged proteins directly from clarified or unclarified cell lysates.

 

HiTrap Columns Prepacked with Sepharose

GSTrap 1ml & 5ml are ready-to-use columns, prepacked with Glutathione Sepharose media.

 

  GSTrap 4B columns are prepacked Glutathione Sepharose 4B columns for convenient, high capacity one-step purification of glutathione S-transferase (GST) tagged proteins.

 

  GSTrap FF columns are prepacked Glutathione Sepharose Fast Flow columns for fast, convenient, one-step purification of glutathione S-transferase (GST) tagged proteins.

 

  GSTrap HP columns are prepacked with Gluthione Sepharose High Performance medium and provide high-resolution, one-step purifications of gluthatione-S-Transferase (GST) tagged proteins.

 

HiPrep Columns Prepacked with Sepharose

GSTPrep is a 20ml ready-to-use column, prepacked with Glutathione Sepharose media.

 

  GSTPrep FF 16/10 is prepacked, ready-to-use column containing Glutathione Sepharose 4 Fast Flow designed for one-step,  scale-up purification of glutathione S-transferase (GST)-tagged proteins.

 

MultiTrap is a ready-to-use 96-well filter plate for high reproducibility.

GST MultiTrap Fast Flow (FF) and GST MultiTrap 4B are prepacked, 96-well filter plates for reproducible, high-throughput screening and rapid parallel purification of gluathione S-transferase (GST) tagged proteins.

 

  GST MultiTrap 4B

 

  GST MultiTrap FF

Sepharose Chromatography Media

Glutathione Sepharose media for purification of GST-tagged proteins.

 

  Glutathione Sepharose 4 Fast Flow is an affinity medium for fast, one-step purification of Glutathione S-Transferase (GST) tagged proteins. Provides good flow properties, excellent for scale up.

 

  Glutathione Sepharose 4B is designed for high capacity single-step purification of Glutathione S-Transferase (GST) tagged fusion.
    Glutathione Sepharose High Performance is an affinity medium for high resolution purification of Glutathione S-transferase (GST) tagged proteins.

 


 GST Buffers

 

  GST Buffers Kit contains prepared buffer concentrates for binding, washing, and elution of GST-tagged proteins.


 GST Detection

For immunodetection of GST-tagged proteins

  Anti-GST Antibody is a polyclonal antibody purified from the sera of goats immunized with purified schistosomal GST.  Because of its polyclonal nature, it can recognize more than one epitope on GST.

 

    GST 96-Well Detection Module permits rapid, sensitive determination of glutathione S-transferase (GST) fusion proteins in a variety of samples.

 

    GST Detection Module is used for identification of GST fusion proteins using either a biochemical or immunological assay.

 


 GST Cleavage

For removal of GST-tags

  Factor Xa, purified from bovine plasma, is a protease used to digest fusion proteins prepared from pGEX vectors containing the recognition sequence for factor Xa (pGEX-3X, pGEX-5X-1, pGEX-5X-2, and pGEX-5X-3). It specifically cleaves following the tetrapeptide Ile-Glu-Gly-Arg.

 

  PreScission Protease is a genetically engineered fusion protein consisting of human rhinovirus 3C protease and GST. It specifically cleaves between the Gln and Gly residues of the recognition seqence of LeuGluValPheGln/GlyPro.

 

  Thrombin is a protease used to digest fusion prepared from pGEX vectors recognition sequence for thrombin (pGEX-1IT, pGEX-2T, pGEX-2TK, pGEX-4T-1, pGEX-4T-2, and pGEX-4T-3. Purified from bovine plasma.

 

 Antibody Purification

Sigma-Aldrich offers GE affinity media for the purification of monoclonal and polyclonal antibodies from different sources. The basis for antibody affinity purification is the high affinity and specificity of protein G and protein A for the Fc region of IgG from a variety of species.

Chromatography media with protein G and protein A immobilized to several different matrices is available, for isolation of IgG and IgG subclasses from ascites, cell culture supernatants, and serum. Chromatography media with recombinant protein L ligand for purification of antibodies and antibody fragments is also available. Protein L has a strong affinity to the variable region of kappa light chains and can be used to capture a wide range of antibody fragments such as Fab fragments, single-chain variable fragments (scFv), and domain antibodies (Dabs).


 Antibody Affinity Chromatography

We offer GE affinity chromatography products for the purification of monoclonal and polyclonal antibodies from different sources. The portfolio includes a range of Sepharose-based chromatography media (resins) using native or recombinant protein A or protein G as ligand. The media are available as lab packs, as well as a variety of column formats, including SpinTrap, MultiTrap, and HiTrap. The MabSelect family allows efficient capture of monoclonal antibodies from large sample volumes. We also offer products for IgM and IgY purification.

Antibody-based products are a growing proportion of new therapeutics on the market and affinity chromatography plays a major role in their production. The basis for antibody affinity chromatography is the high affinity and specificity of protein A and protein G for the Fc-region of IgG from a variety of species.

Protein A and G immobilized on chromatography media and are highly effective in isolating subclasses of IgG from ascites fluid, cell culture supernatants, and serum. Protein A and protein G differ in their ability to bind antibodies of different species and subclasses, and are widely used for the routine purification of antibodies.

IgM Purification

  HiTrap IgM Purification HP is a 1ml column prepacked with Sepahrose High Performance for fast and efficient purification of monoclonal igM.
     • GE17511001

 

  HiTrap IgY Purification HP is a 5 ml column prepacked with Sepharose High Performance for fast and easy purification of IgY from egg yolk.
     • GE17511101

 

Buffer Kits

  Ab Buffer Kit contains carefully prepared buffer concentrates for binding, washing, and elution of IgG according to recommended protocols. The kit also includes neutralizing buffer.
      • GE28903059

 

Capto L

  Capto L is a BioProcess chromatography medium (resin) for the capture of antibodies and antibody fragments. It combines a rigid, high-flow agarose matrix with the immunoglobulin-binding recombinant protein, L ligand, which has a strong affinity to the variable region of antibody kappa light chains. Capto L is suitable for the capture of a wide range of antibody fragments such as Fabs, single-chain variable fragments (scFv), and domain antibodies (Dabs).
     • GE17547806
     • GE17547801
     • GE17547802

 

  HiScreen Capto L columns are prepacked with Capto L, an affinity chromatography medium (resin) for capture of antibodies and antibody fragments such as Fab fragments, scFv, and Dabs. HiScreen Capto L columns are part of the process development platform available from GE Healthcare. The columns are an excellent choice for method optimization and parameter screening.
    • GE17547814

 

  HiTrap Protein L columns are 1 and 5 ml columns prepacked with Capto L, an affinity chromatography medium (resin) for capture of antibodies and antibody fragments such as Fab fragments, scFv, and Dabs.
     • GE29048665
     • GE17547851
     • GE17547815
     • GE17547855

 

GammaBind

    GammaBind G Sepharose® – Recombinant GammaBind is an affinity medium for laboratory scale antibody purification. Binds to all IgG subclasses from human, mouse, and rat; binds total IgG from guinea pig, rabbit, goat, cow, sheep, and horse.
     • GE17088501
     • GE17088502
     • GE17088504

 

    GammaBind Plus Sepharose is used for lab-scale affinity purification of IgG from a large variety of species and has better binding to rat and mouse IgG subclasses compared to GammaBind Sepharose.
     • GE17088601
     • GE17088602
     • GE17088604


 Protein A Antibody Affinity Chromatography

The most widely used affinity chromatography purification step in industry today is the capture of antibodies using Protein A ligand. Our MabSelect family for the capture of monoclonal antibodies (MAbs) has been developed with industrial needs in mind. The expanded product range focuses on improved overall process economy and reduced time for optimization.

We also offer native protein A coupled to Sepharose 4 Fast Flow and Sepharose High Performance chromatography media (resins). Recombinant Protein A is also available coupled to Sepharose-based media.

MabSelect Chromatography Media & Prepacked Columns

Affinity media for capturing MAbs from large sample volumes

HiScreen Columns Prepacked with MabSelect

Excellent choice for method optimization and parameter screening for capture of monoclonal antibodies (MAbs)

 

  HiScreen MabSelect is prepacked with MabSelect and is part of the process development platform available from GE Healthcare. The columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies (MAbs).
     • GE28926973

 

  HiScreen MabSelect SuRe is prepacked with MabSelect SuRe and is part of the process development platform available from GE Healthcare. The columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies (MAbs).
     • GE28926977

 

  HiScreen MabSelect SuRe LX is prepacked with MabSelect SuRe LX and is part of the process development platform available from GE Healthcare. The columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies when a high binding capacity is needed.
     • GE17547415

 

  HiScreen MabSelect Xtra is prepacked with MabSelect Xtra and is part of the process development platform available from GE Healthcare. The columns are an excellent choice for method optimization and parameter screening for capture of monoclonal antibodies (MAbs).
     • GE28926976

 

MabSelect Chromatography Media

Protein A affinity media available in a range of bulk pack sizes.

 

  MabSelect is a protein A affinity medium designed for capturing MAbs from large sample volumes.
     • GE17519901
     • GE17519902

 

  MabSelect SuRe is a protein A affinity medium for capturing MAbs at process scale. MabSelect SuRe has an alkali-tolerant rProtein A ligand that allows the use of rigorous and cost-effective CIP and sanitization protocols based on 0.1 to 0.5 M NaOH. Further, the ligand is protease stable leading to lower ligand leakage.
     • GE17543801
     • GE17543802

 

  MabSelect SuRe LX is a protein A affinity medium with very high dynamic binding capacity at extended residence times and is developed for high titer antibody processes. Alkali tolerance, high capacity and low ligand leakage in combination with the rigid base matric makes MabSelect SuRe LX highly suitable for purification of monoclonal antibodies for clinical applications.
     • GE17547401
     • GE17547402

 

  MabSelect Xtra, with extra high dynamic binding capacity, is a member of the MabSelect family. Developed to meet the demands of increasing levels of expression in monoclonal antibody feedstocks.
     • GE17526907
     • GE17526902

 

HiTrap Columns Prepacked with MabSelect

 

  HiTrap MabSelect columns are prepacked with MabSelect, a Bioprocess medium for capturing of Mabs from large sample volumes.
     • GE28408255
     • GE28408253
     • GE28408256

 

  HiTrap MabSelect SuRe columns are prepacked with MabSelect SuRe, a BioProcess medium for purification of monoclonal antibodies from large sample volumes. The novel alkali-tolerant recombinant Protein A ligand allow cleaning-in-place (CIP).
     • GE29049104
     • GE11003493
     • GE11003494
     • GE11003495

 

  HiTrap MabSelect Xtra columns are prepacked with MabSelect Xtra, a BioProcess medium for capture of monoclonal antibodies from large sample volumes. The increased capacity of the medium is well suited for purification of Mabs from high-level expression feedstock.
     • GE28408258
     • GE28408260
     • GE28408261

 

Protein A Sepharose

Affinity media for purification of monoclonal & polyclonal antibodies.

GraviTrap Columns Prepacked with Protein A Sepharose

rProtein A GraviTrap, and rProtein A Protein G GraviTrap are prepacked gravity-flow columns designed for fast and efficient manual purification of monoclonal and polyclonal antibodies, antibody fragments from cell culture supernatant, and biological fluids.

The antibodies are simply captured with high specificity on protein A gravity-flow columns. You do not need any other instrument with this protocol because the entire process relies on the flow of gravity. The yield varies from 20 to 50 mg of purified antibodies depending on the ligand you use. The columns are reusable up to five times.

GraviTrap Columns are also available prepacked with Protein G Sepharose.

 

  rProtein A GraviTrap includes ten ready-to-use columns prepacked with 1 ml of rProtein A Sepharose Fast Flow.
     • GE28985254

 

  rProtein A Protein G GraviTrap includes ten ready-to-use columns prepacked with 1 ml of a mixture of 50% rProtein A Sepharose Fast Flow and 50% Protein G Sepharose 4 Fast Flow.
     • GE28985256

 

Protein A Sepharose Chromatography Media

 

  nProtein A Sepharose 4 Fast Flow is native protein A coupled to Sepharose 4 Fast Flow for the purification of monoclonal and polyclonal antibodies at both laboratory and process scale.
     • GE17528001
     • GE17528004
     • GE17528002

 

  Protein A Mag Sepharose Xtra are magnetic beads designed to simplify small-scale purification and screening of antibodies. Based on Sepharose with native protein A as ligand.
     • GE28967056
     • GE28967062

 

  Protein A Sepharose CL-4B is a classic, well documented medium for immunoglobulin purification and fractionation, and is suitable for immunoprecipitation procedures.
     • GE17096303
     • GE17096302
     • GE17078001

 

  rmp Protein A Sepharose Fast Flow is a low leakage, non-mammalian based affinity medium designed for high purity separation of monoclonal and polyclonal antibodies at the laboratory and process scales.
     • GE17513801
     • GE17513802
     • GE17513803

 

  rProtein A Sepharose 4 Fast Flow is a well established antibody affinity medium, designed for the purification of monoclonal and polyclonal antibodies at both laboratory and process scale.
     • GE17127901
     • GE17127902
     • GE17127903

 

HiTrap Columns Prepacked with Protein A Sepharose

 

  HiTrap rProtein A FF columns are prepacked with rProtein A Sepharose Fast Flow for purification and fractionation of IgG subclasses and IgG from different sources, including cell supernatants and ascites.
     • GE17507902
     • GE17507901
     • GE28946489
     • GE17508001
     • GE17508002

 

  HiTrap Protein A HP is a prepacked Protein A Sepharose High Performance column for preparative purification of monoclonal and polyclonal antibodies from ascites, serum, and cell culture supernatants.
     • GE17040203
     • GE17040201
     • GE17040301
     • GE17040303


 Protein G Antibody Affinity Chromatography

Affinity chromatography media for IgG purification from a variety of sources. The media is coupled to recombinant protein G from E. coli, with the albumin-binding region of the native protein genetically deleted. Protein G Sepharose 4 Fast Flow and Protein G Sepharose High Performance are available as lab packs as well as prepacked column formats. MabTrap Kit allows fast and simple purification, without the need for a pump, using a prepacked HiTrap Protein G HP column.

Protein G is a bacterial protein and is a good first choice for capture of antibodies at laboratory scale. Compared with protein A, protein G binds a broader range of IgG from eukaryotic species and also binds more IgG subclasses. Typically, protein G has greater affinity for IgG than protein A and binds with minimal affinity to albumin, giving cleaner preparations and greater yields.

The affinity of protein G for IgG depends on the source species and subclass of the immunoglobulin. Besides affinity, the dynamic binding capacity also depends on factors such as flow rate during sample application.

GraviTrap Columns Prepacked with Protein G Sepharose

Protein G GraviTrap and rProtein A Protein G GraviTrap are prepacked gravity-flow columns designed for fast and efficient manual purification of monoclonal and polyclonal antibodies, antibody fragments from cell culture supernatant, and biological fluids.

The antibodies are simply captured with high specificity on protein G gravity-flow columns. You do not need any other instrument with this protocol because the entire process relies on the flow of gravity. The yield varies from 20 to 50 mg of purified antibodies depending on the ligand you use. The columns are reusable up to five times.

GraviTrap Columns are also available prepacked with Protein A Sepharose.

 

  Protein G GraviTrap includes ten ready-to-use columns prepacked with 1 ml of Protein G Sepharose 4 Fast Flow.
      • GE28985255

 

    rProtein A Protein G GraviTrap includes ten ready-to-use columns prepacked with 1 ml of a mixture of 50% rProtein A Sepharose Fast Flow and 50% Protein G Sepharose 4 Fast Flow.
     • GE28985256

 

HiTrap Columns Prepacked with Protein G Sepharose

 

  HiTrap Protein G HP columns are prepacked with Protein G Sepharose High Performance for fast, easy separations of IgG from different species and subclasses.
     • GE17040403
     • GE17040401
     • GE17040501
     • GE17040503

 

    MAbTrap Kit is an affinity chromatography kit, with everything required for small-scale IgG purification from ascites fluid, serum, and cell culture supernatants.
    • GE17112801

SpinTrap Columns Prepacked with Protein G Sepharose

    Ab SpinTrap are prepacked, single-use spin columns for rapid purification of monoclonal and polyclonal antibodies from serum and cell culture supernatants. Designed for small-scale purification of multiple samples in parallel.
     • GE28408347

Protein G Sepharose Chromatography Media

 

  Protein G Mag Sepharose Xtra magnetic beads are designed for rapid, small-scale purification and screening of monoclonal and polyclonal antibodies from serum and cell supernatants. The high capacity, > 27 mg human IgG /ml medium allow for the efficient capture of the antibody with high yield and purity. Approximately 350 μg purified human IgG can be obtained from each purification run.
     • GE28967066
     • GE28967070

 

Protein G Sepharose Chromatography Media

 

  Protein G Sepharose 4 Fast Fllow is recombinant protein G coupled to Sepharose 4 Fast Flow.
     • GE17061801
     • GE17061802
     • GE17061805

 

 Strep-tag™ II-Tagged Protein Purification

Strep-tag™ II is a small tag of only eight amino acids with a relative molecular mass (Mr) of just 1000. Its small size is very beneficial, since in most cases it does not interfere with structural and functional studies, and therefore does not have to be removed.

Strep-tag II binds specifically to StrepTactin™ Sepharose High Performance, which has Strep-Tactin ligand immobilized on a Sepharose base matrix to yield pure target protein. The binding affinity of Strep-tag II to the immobilized ligand is nearly 100-fold greater than to streptavidin, making suitable for purifying Strep-tag II proteins. Purification is run under physiological conditions, and mild elution with desthiobiotin preserves the activity of the target protein.


The small bead size of the Sepharose High Performance chromatography matrix gives high-resolution separations, sharp peaks, and purified target proteins in a concentrated form. StrepTactin™ Sepharose High Performance chromatography medium (resin) is compatible with a wide range of additives, tolerates all commonly used aqueous buffers, and is quickly and easily regenerated using 0.5 M NaOH.

Strep-tag™ II is a small tag of only eight amino acid residues and a molecular weight of just 1000. The small size of the tag is beneficial, since in most cases it does not interfere with structural and functional studies, and therefore does not have to be removed.

Strep-tag™ II binds specifically to StrepTactin™ ligand immobilized on the base matrix to yield pure target protein. The binding affinity of Strep-tag™ to the immobilized ligand is nearly 100-fold greater than to streptavidin, making this method a very efficient way of purifying proteins. Purification is run under physiological conditions, and elution is performed with desthiobiotin, which displaces Strep-tag™ proteins at the biotin-binding site of StrepTactin™. This elution process is mild and the activity of the target protein is thus preserved.


 HiTrap Columns Prepacked with Sepharose

 

  StrepTrap HP is a ready to use HiTrap column prepacked with StrepTactin Sepharose High Performance for purifying Strep-tag II recombinant proteins.
     • GE29048653
     • GE28907546
     • GE28907547
     • GE28907548


 Sepharose Chromatography Media

 

    StrepTactin Sepharose High Performance media for robust, high resolution purification of Strep II-tagged proteins. Mild purification conditions preserve antigenicity and function.
     • GE28935599
     • GE28935600

 MBP-Tagged Protein Purification

Maltose binding protein (MBP) is a useful affinity tag that can increase the expression level and solubility of the resulting tagged protein. The MBP tag also promotes proper folding of the attached protein. Since MBP increases solubility, the tag is particularly useful for recombinant proteins that accumulate in an insoluble form (inclusion bodies).

Affinity purification takes place under physiological conditions and mild elution is performed using maltose. The mild elution preserves the activity of the MBP-tagged protein.


Dextrin Sepharose High Performance chromatography medium (resin) purifies recombinant proteins tagged with maltose binding protein (MBP). The medium is based on small, evenly sized beads that give highly pure products and reduce the need for concentration steps. The medium tolerates commonly used aqueous buffers and is easily regenerated with NaOH. The medium is available in lab packs and prepacked in MBPTrap HP columns.

Maltose-Binding Protein (MBP) is a native E. coli protein and is part of a complex responsible for the uptake and efficient catabolism of maltodextrins. MBP is composed of 370 amino acid residues.

As a fusion tag, MBP is useful for recombinant protein purification by affinity chromatography. It is also used to increase the expression level and solubility of the tagged protein, and for promoting proper folding of the attached protein. MBP increases solubility and the tag is thus particularly useful for recombinant proteins that may otherwise accumulate in an insoluble forms as inclusion bodies. Affinity purification takes place under physiological conditions and mild elution is performed using maltose. The mild elution conditions preserve the activity of the MBP-tagged protein.


 HiTrap Columns Prepacked with Sepharose

 

  BPTrap HP columns prepacked with Dextrin Sepharose High Performance provide convenient, high binding capacity, affinity purification of recombinant proteins tagged with Maltose Binding protein (MBP).
     • GE29048641
     • GE28918778
     • GE28918779
     • GE28918780


 Sepharose Chromatography Media

 

  Dextrin Sepharose High Performance is an affinity media for purification of proteins tagged with Maltose Binding protein (MBP).
     • GE28935597
     • GE28935598