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Brochure: Titan3 and Target2 Chromatography Syringe Filters – Performance Filtration Solutions for Chromatographic Applications Product Literature

Can I filter sterilize Essential 8 Medium using a 0.22 µm filter? Product FAQ

Answer

You can filter the basal Essential 8 medium or the complete Essential 8 medium through a 0.22 µm filter (vacuum driving filtration). But, we recommend that you use a low protein-binding filter.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17399

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Sometimes the ExpiCHO cell supernatant is difficult to filter during clarification. How can I overcome this? Product FAQ

Answer

Because the ExpiCHO system uses components that differ from that of HEK 293-based expression systems such as the Gibco Expi293 system, the supernatant may be more difficult to process through standard bottle top filters in some instances. To remedy this, we recommend centrifuging the supernatant first at ~5,000 x g for 30 minutes followed by filtration using a 0.22 µm filter. Alternative filters (such as depth filters) provide superior filtration for supernatants, especially at larger scales, as these filters are specifically designed to handle supernatant from CHO-derived expressions.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Answer Id: E14746

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How do I use Thermo Scientific Nalgene glass fiber prefilters? Product FAQ

Answer

Glass fiber prefilters are available as an optional accessory (DS0281 series) and bundled with select Thermo Scientific Nalgene Rapid-Flow filter units. They are useful in filtration of solutions with an especially high particulate load, often enabling a higher volume to be filtered before significant flow decay by trapping and preventing the larger particulate matter from entering the final filter membrane. Simply place one prefilter on top of the membrane bonded inside the unit, wet slightly to hold in place, and proceed with filtration as usual.

Answer Id: E15967

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I am using a Nalgene Rapid-Flow Sterile Disposbable Filter Unit and my solution is taking a long time to filter. Do you have any recommendations to improve filtering? Product FAQ

Answer

We recommend using glass fiber prefilters for filtration of solutions with high viscosity or especially high particulate load. The glass fiber prefilters are available as an optional accessory and bundled with select Thermo Scientific Nalgene Rapid-Flow filter units. The prefilters enable a higher volume to be filtered before significant flow decay by trapping and preventing the larger particulate matter from entering the final filter membrane. Simply place one prefilter on top of the membrane bonded inside the unit, wet slightly to hold in place, and proceed with filtration as usual.

Answer Id: E21781

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What type of filter is recommended to prepare liquid aseptic media from Viral Production Medium, AGT powder? Product FAQ

Answer

Viral Production Medium, AGT powder needs to be reconstituted in deionized distilled water or Water For Injection (WFI) for Cell Culture (Cat. No. A12873) and then immediately sterilized by membrane filtration (positive pressure recommended). A low protein binding filter (such as PVDF) with a pore size of 0.2 µM is recommended.

Answer Id: E21201

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How should I filter sterilize CD Hybridoma Medium? Product FAQ

Answer

CD Hybridoma Medium can be filtered by 0.2 µm filtration.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17398

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Do I need to filter the serum before use? Product FAQ

Answer

No, there is no need to filter the serum before use. Gibco serum products are manufactured using the most stringent processes, including membrane filtration.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11867

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Is Gibco Certified Fetal Bovine Serum sterile? Product FAQ

Answer

Fetal bovine serum is triple filter sterilized using a 0.1 µm filter. All our Gibco cell culture liquid products are prepared by an aseptic process for which each step has been validated to ensure that all products meet the industry standard sterility assurance level of 10^-3; i.e., product that demonstrates a contamination level of no more than 1 of 1,000 units during the manufacturing process. The highest level of sterility assurance (equal to or greater than 10^-6) cannot be achieved without terminal sterilization which is harmful to the performance of cell culture products. Key points of control include validated sterilization cycles for all material with product contact, routine media fills employing bacteriological media, a comprehensive environmental monitoring program, validated cleaning procedures, and a validated final filter integrity testing program. Additionally, filtration and dispensing are performed within positive pressure, HEPA-filtered, environmentally controlled rooms.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E19914

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Can I use the SMART Digest Soluble Trypsin Kit with filter-aided sample preparation (FASP)? Product FAQ

Answer

Yes, we recommend the following protocol, which can be optimized as needed:
1. Rinse a Fisher Scientific Vivacon 500, 30kDa MWCO filter (Cat. No. 14558347) to remove trace amounts of glycerin using a fill volume of water. Decant the filtrate and collection vessel. The filter is now ready for use. If you do not want to use the Vivacon filter immediately, then store the pre-rinsed device in the refrigerator with water or buffer covering the membrane surface. Do not allow the membrane to dry out.
2. Dilute sample water to a total volume of 150 µL on the filter.
3. Centrifuge 14,000 RPM for 15 minutes and discard the flow-through.
4. Add 150 µL water.
5. Centrifuge 14,000 RPM for 15 minutes and discard the flow-through.
6. Add 200 µL of SMART Digest buffer.
7. Change the collection vial and add 5 µL of the SMART Digest soluble enzyme to the filter.
8. Parafilm the filters to prevent evaporation of sample during digestion. Incubate the units in a heated bath at 70°C for 60 - 90 minutes.
9. Optional: As needed, add DTT (3 µL of 50 mM DTT), heat at 37°C and incubate for 30 minutes. Alkylate as needed (using 3 µL of 150 mM alkylating reagent).
10. After incubation, add 2 µL of 1% trifluoroacetic acid to the filter. Mix for 1 minute.
11. Centrifuge the filter units at 14,000 RPM for 10 min.
12. As needed, measure protein concentration using a Thermo Scientific NanoDrop Spectrophotometer, or similar procedure.

Answer Id: E20898

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The ProBond resin is not settling down when doing the purification. What could be causing this? Product FAQ

Answer

Chromosomal DNA may be the cause. Ensure that genomic DNA is sheared by pulling the lysate up and down a few times through an 18-gauge needle. Ensure further that the lysate is spun for 15 min at 10,000 rpm in a Sorvall SS-34 rotor in order to clear the lysate. Filtration of the lysate through a 0.8 µm filter is another option.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Answer Id: E12977

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I have sticky cells. What buffer do you recommend to prevent the cells from clumping when using your MagniSort kit? Product FAQ

Answer

For magnetic cell separation, we recommended PBS or HBSS buffers supplemented with 3%-10% fetal bovine serum and 10 mM EDTA. EDTA will help prevent the cells from clumping. Media such as RPMI and DMEM are not recommended because they interfere with the performance of the MagniSort kits. Thorough mixing of the cells and filtration through nylon filters are also recommended to ensure single-cell suspension and removal of debris.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Answer Id: E14631

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A protocol for the use of Nalgene Analytical Filter Funnels and Cellulose Nitrate Membranes in recovery of microorganisms during environmental and/or quality testing Product Literature

How do the Total Exosome Isolation reagents work and what makes them different from the other techniques for exosome isolation? Product FAQ

Answer

Isolation of exosomes is presently a tedious, non-specific, and difficult process. In the course of development of reagents for isolation of exosomes, we evaluated many different technologies, including “gold standard” ultracentrifugation, ultrafiltration; gel-filtration columns, HPLC, and filters. In addition to these simpler methods, we evaluated more advanced approaches including precipitation using various polymers, and bead and column binding using antibodies and various lectins. We also evaluated commercially available products from System Biosciences and other companies. After evaluation, we selected one of the polymers, based on its superior performance, which became the key component of the Total Exosome Isolation reagents (patent application filed). By tying up water molecules, the reagent forces less-soluble components, such as vesicles, out of solution, which allows them to be collected by a short, low-speed centrifugation. The recovered exosomes are then ready for either biological studies or end-point analysis.

Answer Id: E7451

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Legionella and Analytical Filtration Smart Note [DE] Product Literature

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