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GE17-5280-01

nProtein A Sepharose® 4 Fast Flow

Cytiva, 17-5280-01, pack of 5 mL

  •  NACRES NA.56

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Properties

Related Categories Antibodies, Antibody Purification and Characterization, Protein A, G and L Resins, Supplementary Products
shelf life   Please be aware this product may be shipped 90 days before the expiration date. For more information on the batch specific expiration date, please contact technical service.
packaging   pack of 5 mL
mfr. no.   Cytiva, 17-5280-01
matrix   4% cross-linked agarose
average diameter   90 μm (d50v)
cleaning   2 - 10 (pH below 3 is sometimes required to elute strongly bound IgG species. However, protein ligands may hydrolyze at very low pH.)
working range   3 - 9
capacity   >30 mg binding capacity(human IgG/ml)
suitability   suitable for bioprocess medium
storage temp.   2-8°C

Description

General description

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.

nProtein A Sepharose® 4 Fast Flow is native protein A coupled to the well established Sepharose® 4 Fast Flow base matrix. The native protein A ligand is produced by fermenting a selected strain of Staphylococcus aureus. The purified protein is coupled to the cross-linked 4% agarose base matrix by the cyanogen bromide technique, giving a highly stable medium with minimal non-specific adsorption. nProtein A Sepharose® 4 Fast Flow is manufactured without using animal-derived components.

nProtein A Sepharose® 4 Fast Flow has nearly twice the total IgG binding capacity of Protein A Sepharose® CL-4B, and is suitable for recovery and purification of monoclonal antibodies from cell culture at both laboratory and process scale. nProtein A Sepharose® 4 Fast Flow was developed and tested in co-operation with leading manufacturers of purified monoclonal antibody products, and is used in routine commercial production.

As member of the BioProcess media range, nProtein A Sepharose® 4 Fast Flow meets industrial demands with security of supply and comprehensive technical and regulatory support.

Features and Benefits

• Replaces Protein A Sepharose® 4 Fast Flow, the first Cytiva Protein A medium for large-scale purification of antibodies.
• Used in routine commercial production of monoclonal antibodies
• Free from animal-derived components.

Analysis Note

To view the Certificate of Analysis for this product, please visit www.cytiva.com.

Legal Information

Sepharose is a registered trademark of Cytiva

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Column Packing and Preparation for Affinity Chromatography of Antibodies

Use small prepacked columns for chromatography media scouting and method optimization and to increase efficiency in method development.
Keywords: Antimicrobials, Chromatography, Detergents, Gene expression, Separation, Solvents

Desalting and Buffer Exchange for Affinity Chromatography of Antibodies

Desalting at laboratory scale is a well-proven, simple, and fast method that will rapidly remove low molecular weight contaminants at the same time as transferring the sample into the desired buffer ...
Keywords: Absorption, Buffers, Centrifugation, Chromatography, Detergents, Dialysis, Electrophoresis, Fractionation, Gel electrophoresis, High performance liquid chromatography, Liquid chromatography mass spectrometry, Mass spectrometry, PAGE, Precipitation, Sample preparations, Separation, Size-exclusion chromatography, Titrations

Immunoprecipitation Techniques

Target proteins can be isolated and enriched from crude cell lysates by immunoprecipitation (also known as immunoaffinity or pull-down techniques). An antibody selected for its specificity is first affin...
Keywords: Affinity chromatography, Buffers, Cell disruption, Chromatography, Detergents, Electrophoresis, Enzyme activity, Gel electrophoresis, Homogenization, Immobilization, Immunoprecipitation, Mass spectrometry, PAGE, Sample preparations, Separation, Sonication, Western blot

Performing a Purification of IgG Antibodies with Protein G Sepharose® 4 Fast Flow

Protein G Sepharose® 4 Fast Flow consists of 90 µm beads of highly cross-linked agarose, which provide a robust and stable chromatography matrix that allows high flow rates. The medium is a good choi...
Keywords: Buffers, Chromatography, Purification, Sample preparations, Separation

Performing a Purification of IgG Antibodies with nProtein A Sepharose 4 Fast Flow

nProtein A Sepharose® 4 Fast Flow (Figure 3.14) is native protein A coupled to Sepharose® 4 Fast Flow. The medium is designed for recovery and purification of monoclonal antibodies from cell culture s...
Keywords: Buffers, Cell culture, Ligands, Sample preparations

Purification using protein A chromatography media

Protein A is derived from a strain of Staphylococcus aureus and contains five regions that bind to the Fc region of IgG. As an affinity ligand, protein A is coupled to Sepharose® so that these region...
Keywords: Affinity chromatography, Buffers, Chromatography, Fermentation, Immunoprecipitation, Ligands, Purification, Size-exclusion chromatography

Protocols

Affinity Chromatography Troubleshooting

This section focuses on practical problems that may occur when running a chromatography column. The diagrams below give an indication of how a chromatogram may deviate from the ideal during affnity p...
Keywords: Adsorption, Affinity chromatography, Buffers, Chromatography, Detergents, Digestions, Fractionation, Ligands, PAGE, Precipitation, Purification, Sample preparations, Separation, Solvents

Buffer Exchange and Desalting for Affinity Chromatography

Dialysis is frequently mentioned in the literature as a technique to remove salt or other small molecules and to exchange the buffer composition of a sample. However, dialysis is generally a very slo...
Keywords: Absorption, Buffers, Dialysis, Separation

Column Packing and Preparation for Affinity Chromatography

Prepacked columns from Cytiva will ensure reproducible results and the highest performance. However, if column packing is required, the following guidelines will apply at any scale of operation:
Keywords: Antimicrobials, Buffers, Separation

Converting From Linear Fow (cm/hour) to Volumetric Flow Rates (ml/min) and Vice Versa

It is convenient when comparing results for columns of different sizes to express flow as linear flow (cm/hour). However, flow is usually measured in volumetric flow rate (ml/min).

Performing a Separation with Cytiva Products Based on Protein A

Protein A is derived from a strain of Staphylococcus aureus and contains five regions that bind to the Fc region of IgG. As an affinity ligand, protein A is coupled to Sepharose so that these regions a...
Keywords: Affinity chromatography, Buffers, Cell culture, Chromatography, Ligands, PAGE, Purification

Pull-down assays

Pull-down assays involve isolation of a protein complex by adsorbing the complex onto beads. Immobilized ligands on the beads bind specifically to a component of the complex, either via an affinity tag...
Keywords: Buffers, Cell disruption, Centrifugation, Chromatography, Cloning, Digestions, Gene expression, Genetic, Immobilization, Immunoprecipitation, Ligands, Mass spectrometry, PAGE, Precipitation

Sample Preparation for Affinity Chromatography

Samples for chromatographic purifcation should be clear and free from particulate matter. Simple steps to clarify a sample before beginning purifcation will avoid clogging the column, may reduce the ...
Keywords: Adsorption, Affinity chromatography, Buffers, Centrifugation, Chromatography, Detergents, Filtration, Nucleic acid denaturation, PAGE, Precipitation, Sample preparations

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