Product FAQ

Do you offer magnetic bead-based kits for avidin-biotin affinity purification?

Answer

Please refer to Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support for information available on the following link (https://www.thermofisher.com/us/en/home/technical-resources/technical-reference-library/protein-purification-isolation-support-center/protein-ip-coip-pulldown-support.html).

Answer Id: E12909

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

How does the NativePure™ Affinity Purification System work?

Answer

pcDNA™ vectors with “capTEV™” tags (BioEase™ in vivo biotinylation peptide + 2 TEV protease recognition sites + 6X His tag) allow expression of biotinylated recombinant fusion proteins in mammalian cells. The protein complexes are purified under native conditions using streptavidin agarose included in the kit.

Answer Id: E12910

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What are the specifications of the NativePure™ Columns?

Answer

The NativePure™ Columns hold up to 2 mL of chromatography resin and contain a 10 mL reservoir for sample or buffer. The column specifications are listed below:

-Pore size of purification columns: 30-35 microns
-Recommended flow rate: 0.5 mL/min
-Maximum flow rate: 2 mL/min
-Maximum linear flow rate: 700 cm/h
-Column material: polypropylene
-pH stability (long term): pH 3-13
-pH stability (short term): pH 2-14

Answer Id: E12911

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What kind of tubes can I use to form DNA:lipid complexes?

Answer

Polypropylene, polystyrene, or glass tubes may be used with any of our transfection products.

Answer Id: E3124

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

CaptureSelect™ Affinity Purification What are the benefits of CaptureSelect™ affinity products?

Answer

CaptureSelect™ affinity resins can make the purification of biomolecules that do not have a traditional affinity purification solution much more efficient. When a targeted, specific affinity purification solution does not exist for a biomolecule, the protein purification scheme can be very complex needing 4-5 chromatography steps. Many chromatographic steps need to be utilized to separate the product of interest from key process and product related impurities. As the number of required purification unit operations increases, the product yield decreases and is heavily impacted with each step added. Yield drives cost of goods, so not having an affinity purification solution can greatly impact the cost for biotherapeutic manufacturing. Using in-house capabilities and expertise, CaptureSelect™ Custom Services design product-specific affinity ligand based on single-chain, camelid-derived antibodies, and couple that ligand to a high-performance, cost-effective affinity resin. We currently offer off-the-shelf bioprocess resins for antibody fragments, biosimilars, and viral vectors. Custom ligands can be used for biomolecule purification, scavenging of challenging impurities, and quantitation or detection of biomolecules. Custom resins are suitable for use in a cGMP production process.

Answer Id: E12912

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Do you offer a kit for antibody purification?

Answer

For purification of antibodies, we offer Melon™ Gel, which binds other proteins from serum, ascites or culture supernatant, allowing the IgG to be collected in the flow-through. As low pH is typically used to elute IgG from Protein A, G, A/G or L, the use of Melon™ Gel can spare the IgG from exposure to such harsh conditions, which can be particularly important for monoclonal antibodies.

Answer Id: E12929

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

I am getting a “Change in Constant Mode” error on my PowerEase™ 90W Power Supply? Why is this?

Answer

Here are the scenarios for this error with likely causes and solutions:

- Voltage changes to current. Current limit was set too low due to which the ceiling for current was hit and constant mode changed from voltage to current. Increase current limit to 500 mA.
- Current changes to voltage. Voltage was set to low due to which the ceiling for voltage was hit and constant mode changed from current to voltage. Increase voltage limit to 300 V.

Answer Id: E11024

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

I have optimized my conditions for a particular cell line but I seem to be getting inconsistent results with my transfection. What factors contribute to this inconsistency?

Answer

Cells from a different passage number may behave differently. Also, if cells were sitting at confluence prior to plating for transfection, they may not transfect efficiently. To minimize such inconsistencies, passage the cells while they are still growing exponentially. Actively dividing cells transfect better.

Answer Id: E3125

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What are the benefits of POROS™ chromatography resins in a manufacturing process?

Answer

Increased titers and product demand have caused substantial bottlenecks in downstream processing for a range of biomolecules. POROS™ chromatography resins address these challenges with solutions that help maintain performance and add process flexibility. Due to the polymeric nature of the backbone and the way we attach the ligands, the beads have very robust physical and chemical stability. This allows for aggressive solutions to be used, if needed, to improve cleanability and thereby increase resin lifetime/reuse. Our ion exchange resins are the go-to resins in downstream monoclonal antibody and recombinant protein chromatography where capacity, resolution, and yield are critical. We are also working on a new line of hydrophobic interaction resins that may be of interest for future projects that will be a best-in-class product for capacity and resolution with an added benefit of being able to load under lower salt concentrations and faster flow rates. We are also a strong partner when it comes to quality and supply of our products.

Answer Id: E12913

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What are fusion tags?

Answer

Fusion tags are pieces of proteins (such as glutathione S-transferase (GST)) or amino acid sequences (such as His-Tag, c- Myc tag, or HA tag) that can be added to a protein expressed in a cultured cell in vitro and can be easily detected or purified. The resulting proteins are referred to as recombinant proteins as the DNA for the original protein has been recombined with the DNA of the fusion tag in order to produce one protein that has sequences of both the original protein and the fusion tag. Adding a tag to a protein can give it a specific binding affinity.

Fusion proteins (recombinant proteins with fusion tags) can be produced (expressed) in prokaryotic cells, such as E. coli or eukaryotic cells, such as mammalian cells. Fusion proteins expressed in eukaryotes may be glycosylated or otherwise posttranslationally modified. The systems for these modifications are typically missing in prokaryotic cells.

Answer Id: E12930

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

The fuse on my PowerEase™ 90W Power Supply has blown out? How do I replace it?

Answer

One extra fuse is supplied with the PowerEase™ 90W Power Supply. For detailed instructions on replacing the fuse, please refer to Page 17 of the manual.

Answer Id: E11025

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Can I use the same amount of any lipid reagent for transfection of my particular cell line?

Answer

No. The amount of lipid for each lipid reagent should be optimized for each cell line. Each lipid reagent has different composition/formulation, which will have different impact on each cell line. Therefore please optimize whenever you have a new lipid for each cell line.

Answer Id: E3126

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What type of column do you recommend for POROS™ 50 μm resins? Do I need a high pressure column for POROS™ resins?

Answer

POROS™ resins are mechanically rigid and incompressible and can be packed effectively in low-pressure glass columns and in high-pressure stainless steel columns. The lack of wall support with increasing column diameter has minimal impact on chromatography performance because the beads support themselves, allowing for flexible column packing approaches and consistent and robust results. Columns can be packed with traditional flow pack, axial compression, or pack-in-place/stall pack packing methods.

Answer Id: E12914

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What type of recombinant protein purification resins do you offer?

Answer

We offer a variety of resins targeting His-tagged fusion proteins or GST fusion proteins. Please see this selection table to choose the right resin for your experiments. Table available at http://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-purification-isolation/protein-purification/recombinant-protein-purification.html

We also offer resins (both agarose and magnetic bead-based) with immobilized anti-c-Myc and anti-HA antibodies for targeting c-Myc and HA-tagged proteins respectively. Please check out the following links for the specific products:

-Anti-c-Myc agarose (https://www.thermofisher.com/order/catalog/product/20169?ICID=search-product)
-Anti-c Myc magnetic beads (https://www.thermofisher.com/order/catalog/product/88843?ICID=search-product)
-Anti-HA agarose (https://www.thermofisher.com/order/genome-database/antibody/HA-Epitope-Tag-Antibody-clone-2-2-2-14-Monoclonal/26181)
-Anti-HA magnetic beads: (https://www.thermofisher.com/order/catalog/product/88837?ICID=search-product)

Answer Id: E12931

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What alternative do you offer for the discontinued SensiCell™ MEM?

Answer

The alternative product is Advanced MEM (Cat. No. 12492-013). Please add 2 mM GlutaMAX™ Supplement to the medium.

Answer Id: E13255

Was this answer helpful?

Yes
No
Thank you for your response