:: Posted by American Biotechnologist on 04-22-2010
Protein overexpression and affinity purification are used in various biotechnology research fields and play an important role in the post genomic era. While many of the common purification techniques make use of tags like 6XHis and GST, they face drawbacks such as significant background and an extra tag-removal step (especially if the tag is known to interfere with the protein’s function).
Bio-Rad’s Profinity eXact protein purification system utilizes an immobilized engineered protease that specifically recognizes and binds the Profinity eXact tag with high affinity. Following purification, the protein is washed and the tag is cleaved while still on-column.
This technical note demonstrates how the Profinity eXact purification system overcomes concerns common to expression and purification systems in eukaryotic cells including: poor tag expression, tag degradation, posttranslational modification and misfolding of tag and excessive background contamination. The results indicate that the Profinity eXact system can be used in eukaryotic cells for single-step purification of tag-free proteins with low background and without compromising the obvious function of the system.
Faster, Cleaner, More Specific Affinity Tag Purification
:: Posted by American Biotechnologist on 04-01-2010
As discussed in an earlier post on siRNA delivery, Bio-Rad’s Gene Pulser MXCell Electroporator is a powerful tool for biotechnology labs involved in delivering siRNA into difficult to transfect cells. In this technical note, biotechnology experts from the University of Rochester demonstrate how the MXCell can be used to simultaneously optimize electroporation conditions in multiple cell types thereby saving time, reagents and effort.
Electroporation of Smooth Muscle Cells Using the Gene Pulser MXcell Electroporation System
:: Posted by American Biotechnologist on 03-31-2010
Small interfereing RNA (siRNA) technology is a powerful tool for facilitating post-transcriptional gene silencing. As with most transfection techniques, siRNAs have been traditionally delivered into the cell by one of four transfection techniques: lipid-mediated, chemical-mediated, viral transfection or electroporation.
In this paper, a team from the Beckman Research Institue of the City of Hope demonstrate how Bio-Rad’s MXCell electroporator can be used to optimize transfection conditions for B-Cell lymphomas which are extremely difficlut to transfect by most traditional methods.
Delivering siRNA into Difficult to Transfect Cells Using Bio-Rad’s Gene Pulser MXcell Electroporation System
:: Posted by American Biotechnologist on 03-25-2010
The Bio-Rad ProteOn XPR36 is a surface plasmon resonance (SPR) system capable of analyzing 36 protein-protein interactions in one experiment. Up to 6 antibodies may be simultaneously immobilized in parallel channels and assayed with panels of up to 6 antigens.
The Antibody Characterization and Development Using the Bio-Rad ProteOn technical note describes how the ProteOn can be used for:
Hybridoma screening and ranking
Kinetic analysis of antibody-antigen specificity
Determination of cross-reactivity of antibodies used in multiplex assays
For a great primer on Bio-Rad’s ProteOn XPR36 check out this brief video tour
:: Posted by American Biotechnologist on 03-18-2010
Affinity purification of protein is a common technique in most protein labs which is often the bottle neck in achieving faster experimental results. The technique usually involves the use of HIS or GST tags which need to be enzymatically cleaved from of the recombinant protein in a laborious and time-consuming process so as not to interfere with the protein’s functional activity or crystallization process. Furthermore, many functional assays and crystallography experiments require large amounts of protein which can be frustrated by the drawn-out purification/tag cleavage process.
In this technical note, Justin L. Burns et. al. from the Georga Insitute of Technology report how the combination of the Profinity eXact fusion-tag and Profinia automated purification system allowed them to recover milligram quantities of protein from a gram-negative bacteria in under 2 hours.
Purify Protein in Under 120 Minutes