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Guide to handling and storage of prepared plate media Product Literature

Product information: Guide to handling and storage of prepared plate media Product Literature

Catalog: European Prepared Media Selection Guide Product Literature

How flexible is the Gibco Essential 8 Flex Medium feeding schedule? Can you give an example feeding schedule? Product FAQ

Answer

Best results are observed when cells are split twice weekly. To enable a weekend-free culture schedule, cells would be split on Monday and Thursday with full media exchanges on Tuesday and Friday, approximately 24 hours after plating. In this example, the Friday feed should use twice the standard volume.

Answer Id:: E10272

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Non Hazardous Tubed - Bottled Prepared Media [NZ] SDS

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Non Hazardous Tubed - Bottled Prepared Media [AUS] SDS

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Are cell density (% confluency) and passage number important considerations for transfection? Product FAQ

Answer

Yes, cell density is an important parameter in influencing transfection efficiency. If the seeding density is too low, some cytoxicity may be observed. If the cell density is high, lower than expected transfection efficiency may be observed. Both issues may be easily resolved by either descreasing or increasing the quantity of complexes added to the culture. We recommend using Lipofectamine 3000 since it shows the best flexibility for variable seeding density without showing cytotoxicity issues and maintains high protein expression. Lipofectamine 3000, Lipofectamine 2000, and Lipofectamine LTX/PLUS provide excellent transfection efficiencies at confluencies between 70 and 90%. Some toxicity may be observed at lower confluencies but may be alleviated by decreasing quantity of complexes or removing the complexes after 4-6 hours incubation and refreshing the media. Lipofectamine RNAiMAX works best at confluencies between 60 and 80%.

Passage number may affect transfection experiments. We recommend consistent splitting and plating of cells. Excessive numbers of passages may decrease transfection performance. We do not recommend splitting cells for more than 20-30 passages. If transfection performance declines and cells have been in culture for a long time or excessively/improperly passaged, we recommend that you restart your cultures with a new vial of cells from liquid nitrogen. Please refer to the Gibco Cell Culture Basics handbook (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics.html) for proper guidelines for culturing and passaging cells.

Answer Id:: E8962

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Streamline Your Media Preparation Product Literature

I need to buy media to prepare agar plates, which I will be using to grow colonies after transformation (molecular cloning). Which product would you recommend? Product FAQ

Answer

We offer two options:

The first is our formulation of LB agar (catalog number 22700-025), which has LB Media and agar mixed together. This comes in a 500g and 2.5 kg size. All you have to do is measure out, add water, and autoclave.

The second option is even easier. Our imMedia comes premeasured in pouches. We have three types: imMedia Amp Agar, Kan Agar and Zeo Agar for ampicillin, kanamycin and Zeocin-containing plates. All you have to do is empty the pouch, add water, heat and pour plates. No autoclaving is needed! Each pouch contains sufficient reagents for 8-10 plates.

imMedia Amp Agar, 20 pouches, Q601-20

imMedia Kan Agar, 20 pouches, Q611-20

imMedia Zeo Agar, 20 pouches, Q621-20

For added convenience, you should also check out our LB medium imMedia products: (Each pouch makes 200 mL of media)

imMedia Amp Liquid, 20 pouches, Q600-20

imMedia Kan Liquid, 20 pouches, Q610-20

imMedia Zeo Liquid, 20 pouches, Q620-20

Answer Id:: E3039

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What is the Beta Amyloid ELISA SHSY5Y cell supernatant sample preparation protocol? Product FAQ

Answer

Preparation of SHSY5Y cell supernatant samples for testing with Beta Amyloid ELISA:
1) Plate SHSY5Y cells (neuroblastoma) so that 90% confluency will be reached on the second day.
2) Wash cells 1X with full media (DMEM, 10% FBS, with phenol red or without).
3) Cover cells with full media (~550 microliters).
4) Incubate 36 to 40 hours. If media becomes acid and turns a strong yellow color, beta amyloid can degrade.
5) Add PMSF to collection tubes (500X, 20 mg/mL in dry isopropanol). Mix gently and centrifuge out cells.
6) Stabilize samples by mixing (10:1) with IP Buffer containing protease inhibitors. The 10X IP Buffer is: 500 mM Tris, pH 7.4, 1.5 M NP-40, 5% deoxycholate.
7) If cell line expresses exogenous APP, dilute samples with DMEM/IP Buffer with inhibitors.
8) Run samples in ELISA as recommended in product insert.
9) Preparing standard curve dilutions in DMEM/IP Buffer with inhibitors is recommended to provide the same matrix as the samples.

Answer Id:: E5198

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Guidance for configuring your automated plate processing system Product Literature

Lot # 4341584 Certificate

Catalog # PP2010

Lot # 4338704 Certificate

Catalog # PP2010

Lot # 4344978 Certificate

Catalog # PP2010

Lot # 4339693 Certificate

Catalog # PP2010
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