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Why do I need to pre-rinse all materials that will come in contact with primary neurons/astrocytes with complete medium? Product FAQ

Answer

These cells readily stick to the plastic used in cell culture dishes and centrifuge tubes. Prior to use, rinse all materials that will come in contact with the cells with medium to prevent cells from sticking to the plastic.

Answer Id: E12009

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Nunc Skin Graft Cell Culture Dish Product Literature

Specificiation Sheet: Thermo Scientific Nunc EasYDish cell culture dishes Product Literature

I would like to use Collagen I Rat Protein, Tail (Cat. No. A1048301) for coating cell culture dishes for growth of cardiac fibroblasts. Can you please provide a coating protocol? Product FAQ

Answer

For your application, the thin coating procedure should be fine at a starting concentration of 5 µg/cm2. You can dilute the collagen in 20 mM acetic acid to the volume needed. Coat the surface, incubate 1 hour at room temp, aspirate the solution, rinse 3 times with PBS, and use or store plates.

See the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/Collagen_I_rat_tail_PI.pdf) for details.

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

Answer Id: E17376

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I'm performing an intracellular crosslinking experiment with BS3 on adherent cells. When quenching, do I need to aspirate the BS3 first and then add the quench solution to my cells? Is there a recommended volume of cross-linker and quench solution to add? Product FAQ

Answer

BS3 is a water-soluble crosslinker but it is not membrane permeable. DSS is the membrane-permeable alternative if the aim is to perform intracellular crosslinking. Either the BS3 or DSS solution should be added at a final concentration of 1-5 mM. Sufficient volume used should be enough to cover the surface of the cells: see the link below for volumes typically used for cell culture media. For example, use about 2 mL of media for a 6-well plate well. Use the same volume for the crosslinker. Refer to our chart (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html) for useful information of various sizes of cell culture dishes and flasks.
After incubating the crosslinker solution, you do not need to aspirate it. Instead add the Tris quenching buffer to a final concentration of 10–20 mM Tris. Based on the stock concentration of your Tris quenching buffer, you can determine the volume need to get the 10–20 mM final concentration.

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

Answer Id: E15729

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I would like to use Collagen I Rat Protein, Tail (Cat. No. A1048301) for coating cell culture dishes. In the first step of the thin coating protocol, what should I use to dilute the 17.4 M acetic acid stock solution to 20 mM? Product FAQ

Answer

You can just use water to dilute the acetic acid stock solution to 20 mM.

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

Answer Id: E17377

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What is the recommended working volume of Nunc adherent cell culture products? Product FAQ

Answer

This depends largely on the cell line and working conditions (considerations include headspace for gas exchange, supplements used, and optimal feeding schedule, among others) but we do offer guidelines for development purposes (0.3 to 0.5 mL per cm2) . Note that these volumes are approximate and may have to be modified for best performance. T25 Flask = 7 mL, T75 Flask = 25 mL, T175 Flask = 55 mL, T225 Flask = 70 mL, 300cm2 Flask = 100 mL, 500 cm2 TripleFlask = 200 mL, 35 mm round culture dish = 3 mL, 60 mm round culture dish = 5 mL, 100 mm round culture dish = 12 mL, 150 mm round culture dish = 35 mL, 4 (rectangular) well Multidish = 5 mL, 8 (rectangular) well Multidish = 3 mL, 4 (round) well Multidish = 1 mL, 6 well Multidish = 3 mL, 12 well Multidish = 2 mL, 24 well Multidish = 1 mL, 48 well Multidish = 0.5 mL.

Answer Id: E15931

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细胞培养的技巧及相关产品选择 Product Literature

Which format of cultures should I use for neural induction with PSC Neural Induction Medium? Product FAQ

Answer

Neural induction can be started with human PSCs cultured on 6-well plate or culture dishes. PSCs cultured in flasks are not recommended because it is difficult to remove non-neural differentiated colonies in flasks.

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

Answer Id: E20499

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What are mesenchymal stem cells? Product FAQ

Answer

Mesenchymal stem cells (MSCs) are multipotent cells isolated primarily from bone marrow or fat tissues that exhibit the ability to differentiate into bone, cartilage, and fat cells. Under normal cell culture conditions, MSCs isolated from bone marrow are spindle shaped with the unique ability to adhere to uncoated plastic culture dishes (Arthritis Res Ther 9:204 (2007)).

Answer Id: E12184

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What is Nunclon Delta? Product FAQ

Answer

Nunclon Delta refers to our performance-certified surface for culture of adherent cells, available in a variety of common formats (flasks, plates, multiwell dishes). It is tested with at least four cell lines (varies by product and manufacturing location) commonly used in research, and it should work well for most robust cell lines. It does not involve addition of coating reagent, but rather is manufactured by exposure of the molded products to a proprietary energy source, which renders the surface more hydrophilic by means of increasing the concentration of oxygen-containing (e.g. hydroxyl) molecular groups. It is by this hydrophilic property that anchorage-dependent cells are able to attach to the surface and become confluent.

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

Answer Id: E15927

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With the Nunc Polycarbonate Cell Culture Inserts in Multi-Well Plates, what is the distance between the cell culture membrane and the bottom of the multi-well dish? Product FAQ

Answer

The membrane is approximately 1.0 mm above the bottom of the dish for the Nunc Polycarbonate Cell Culture Inserts.

Answer Id: E17498

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Technical Bulletin: Nunc Nunclon TripleFlask Culturing Technique Product Literature

Is it normal to see floating debris when pre-coating with Ready-to-Use Gibco Geltrex Matrix? Product FAQ

Answer

Yes, this is normal. It has been observed in product development. Remove the Gibco Geltrex Matrix solution after pre-coating and add cells as normal. H9 human embryonic stem cells have been cultured on dishes pre-coated with Ready-to-Use Gibco Geltrex Matrix with this observation in the pre-coating procedure.

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

Answer Id: E11782

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Is it normal to see floating debris when pre-coating with Ready-to-Use Geltrex Matrix? Product FAQ

Answer

Yes, this is normal. It has been observed in product development. Remove the Geltrex solution after pre-coating and add cells as normal. H9 human embryonic stem cells have been cultured on dishes pre-coated with Ready-to-Use Geltrex Matrix with this observation in the pre-coating procedure.

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

Answer Id: E11817

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