Biotinylated Protein Purification

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Brochure: Refine Protein Preparation
Purification or Removal of Biotin and Biotinylated Substances

Agarose beads and affinity gels

Agarose beads and affinity gels enable biotinylated protein and biotinylated antibody purification using streptavidin or avidin cross-linked to beaded agarose. Both avidin and streptavidin bind to biotin with high affinity, though avidin can exhibit high non-specific binding. Extravidin®-agarose uses a modified avidin with very high affinity and specificity for biotin. Good for small to large scale purification, these resins are suitable for gravity flow column purification, low-speed centrifugation in batch processing, and low-pressure chromatography procedures. EZview™ affinity gels incorporate a red dye that allows the user to easily differentiate pellet from supernatant, resulting in less protein loss during batch purification.

Magnetic beads

PureProteome™ Streptavidin magnetic beads provide a rapid and reproducible way to purify immunoglobulins from complex mixtures such as serum, plasma or cell culture supernatant samples. Biotinylated proteins and biotinylated antibodies are purified using a magnetic rack or platform to separate beads from wash and elution fractions. This procedure is suitable for low throughput using single microfuge tubes or higher throughput using 96-well plates with or without automation. Streptavidin Magnetic Particles can also be used for purification of biotinylated proteins and biotinylated antibodies.

SigmaScreen™ Streptavidin High Capacity Coated Plates

SigmaScreen™ streptavidin-coated high capacity plates utilize a proprietary coating technology that provides substantially greater biotin binding capacity than standard streptavidin or ExtrAvidin coated plates. With binding capacity >300 pmol/well (96-well format) or >150 pmol/well (384-well format), these plates allow the captured protein to be eluted for post-capture analysis by various methods such as MALDI, ICAT or SDS-PAGE. SigmaScreen™ streptavidin-coated high capacity plates are manufactured under ISO 9002 in our GMP facility and deliver high well-to-well and plate-to-plate consistency.

Cytiva™ Amersham chromatography columns and media

A variety of Cytiva™ Amersham chromatography columns and media resins are available for reliable, high resolution small- to large-scale purification of biotinylated proteins and antibodies.

SpinTrap™ columns are prepacked, single-use spin columns for simple, small-scale purification of biotinylated proteins and biotinylated antibodies.
HiTrap™ HP columns are prepacked with Streptavidin Sepharose™ High Performance resin for routine, preparative purification of biotinylated proteins and antibodies.
MultiTrap™ 96-well filter plates provide a fast, reliable format for small-scale purification of biotinylated proteins and antibodies using prepacked 96-well filter plates amenable to centrifugation or vacuum filtration, manually or using automated robotic systems.
Streptavidin Mag Sepharose™ media are comprised of Sepharose™-based magnetic beads designed to simplify enrichment of target proteins by immunoprecipitation or pull-down applications, and can be used for rapid, small-scale purification and screening of biotinylated proteins and biotinylated antibodies from cell supernatants.
Streptavidin Sepharose™ High Performance resin is also available separately for scaling purification.

Related Applications

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Protein Expression

Protein expression technologies for expressing recombinant proteins in E. coli, insect, yeast, and mammalian expression systems for fundamental research and the support of therapeutics and vaccine production.

Lysis & Protein Extraction

An overview of cell lysis and protein extraction methods including detergent solubilization, freeze-thaw lysis, osmotic shock, sonication, enzymatic cell lysis, and mechanical disruption techniques such as Dounce, Polytron, and mortar and pestle homogenization.

Protein Pull-Down

Pull-down assays, reagents, and protocols for investigating in vitro protein-protein interactions using affinity or GST pull-down, tandem affinity purification (TAP), and co-immunoprecipitation methods.

Protein Concentration & Buffer Exchange

Overview of techniques used to concentrate and clarify protein samples for purification, bioprocessing, and analysis workflows. Includes protocols and videos for filtration and ultrafiltration techniques, protein enrichment, and desalting and buffer exchange using dialysis, diafiltration, and chromatography methods.

Gel Electrophoresis

Protein gel electrophoresis is used to separate proteins for purification, characterization, and detection. Methods such as native PAGE, SDS-PAGE, 2D PAGE, and isoelectric focusing (IEF) are used in preparation for downstream applications including Western blotting, mass spectrometry, and proteomic analysis.

Western Blotting

An overview of the principles and applications of Western blotting for protein detection. Includes links to detailed protocols, videos, and other resources for protein extraction, gel electrophoresis, transfer to PVDF or nitrocellulose membranes, and chemiluminescent, colorimetric, and fluorescent detection methods.

Mass Spectrometry

Mass spectrometry (MS) is an analytical tool used to identify compounds, determine chemical structure, and assess isotopic abundance. In MS, samples are ionized, and the resulting ions are identified based on their mass-to-charge (m/z) ratios.