Streptavidin Columns

The challenge of modern Target Discovery requires the efficient automation of complex assays. The immunoprecipitation (IP) assay has proven quite useful in research discovery to identify components of molecular complexes, discovering protein-protein and protein-nucleic acid interactions, and quantifying an immunogenic response.

Protein A PhyTip® columns have been successfully used for this purpose, but IP can only be successful if an antibody is available to the peptide or protein of interest. The availability of biotin labeling reagents allows researchers to generate customized affinity resins by conjugating proteins of interest to streptavidin resins. These custom resins are then ready to pull down interactive proteins and complexes from biological samples. To efficiently automate these assays, significant sample preparation steps must be addressed. Namely, protein needs to be purified, excess biotin labeling reagent must be removed, and biotinylated protein needs to be immobilized.

PhyNexus Affinity Purification, Gel Filtration columns containing resin of 5K molecular weight cutoff, and Streptavidin columns in combination are the ideal tools for small-volume, automated, high throughput purification, separation, and immobilization of biotinylated proteins for complex assays.

Description

PhyTip columns are unique capture, purification and enrichment™ tools from PhyNexus designed for lab-scale protein sample preparation. PhyTip columns are available for a variety of liquid handling platforms and contain specific affinity resins for application specific requirements.

PhyNexus recommends the use of biotin-labeled bovine albumin (Sigma, A8549) as a standard to verify the use of PhyTip columns with all applications.

Samples for purification and enrichment must be clear and free from particulate matter. It is highly recommended to centrifuge samples and use the clear supernatant only, prior to use with PhyTip columns.

Application

PhyTip columns with Streptavidin PhyTip columns with Streptavidin have been optimized for use with general molecular biology laboratory reagents and specific PhyNexus instrument flow rates/volumes as shown below. This information was collected using the PhyNexus Purification Systems.

PhyNexus recommends the following buffers (not supplied):

Capture Buffer – Phosphate Buffer solution pH 7.4, for those situations where additional buffer needs to be added to supplement the volume of the sample and to ensure correct pH for capture

Wash Buffer I – Phosphate Buffer solution pH 7.4

Wash Buffer II – Phosphate Buffer solution pH 7.4

Enrichment Buffer – If performing a pull-down assay, for the final elution step may be compatible with an acidic buffer such as Phosphate Buffer solution pH 2.5

Neutralization Buffer. – Tris Buffer solution pH 9.0

Data

PhyNexus recommends the use of the PhyTip Starter Kit containing IgG as a standard to verify the use of PhyTip columns with all applications.

Samples for purification and enrichment must be clear and free from particulate matter. It is highly recommended to centrifuge samples and use the clear supernatant only, prior to use with PhyTip columns.

Shipping and Storage

Each pack of PhyTip columns has been manufactured and QC’d to the highest standards and shipped in retainer boxes that maintain the integrity of the specific affinity resin within each PhyTip column. This product is shipped at ambient temperatures, but on receipt should be stored in a standard laboratory refrigerator between 4 and 8◦C.

  • Do NOT freeze or store frozen.
  • When not in use, keep the lid of the box closed and sealed, store in the refrigerator.
  • Do not allow affinity resin to dry out by extended storage in a dry environment.

PhyTip columns with Streptavidin are stored in Glycerol when shipped from PhyNexus.

Documents
Robotics Systems Compatibility

Biotin-tagged PhyTip Columns are available for these robotic systems.

Part Numbers

Streptavidin PhyTip Columns