What reagents should I use to stain my protein gels?

Product FAQ

Answer

Any dye in the 625 nm emission range should work. For example, you can use the WesternDot™ 625 Western Blot Kit. We also offer our White-Light Conversion Screen (Cat. No. 4473061), which converts blue light emitted by the blue-light transilluminator or UV light emitted by the UV-light transilluminator to white light. This conversion screen is compatible with multiple protein stains including SimplyBlue™ SafeStain, SilverQuest™ silver, and Coomassie™ blue stains.

Answer Id: E7990

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Are there robotic scripts for running E-Gel™ 48/96 or E-PAGE™ 48/96 gels?

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Answer

Robotic scripts for running E-Gel™ 96 Agarose Gels and E-PAGE™ 96 Gels on Beckman Coulter's Biomek™ FX workstation can be found on our website at: www.thermofisher.com/egels (click the Labware Definitions link in the left navigation pane).

Answer Id: E7991

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Do you have the recipe for the TBE running buffer and the Hi-Density TBE Sample Buffer?

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Answer

Please see the recipes below:

TBE Running Buffer (5X):
Tris base, 54.0 g
Boric acid, 27.5 g
EDTA (free acid), 2.9 g
Deionized water to 1.0 L

Hi-Density TBE Sample Buffer (5X):
5X TBE Running buffer (Cat. No. LC6675), 2 mL
Ficoll™ Type 400, 1.5 g
Bromophenol blue, 1 mL of 1% solution
Xylene cyanol 1 mL of 1% solution
Deionized water to 10.0 mL

Answer Id: E7992

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What are the shelf life and storage conditions for my TBE gels?

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Answer

The shelf life is 8 weeks for the gels, which should be stored at 4°C.

Answer Id: E7993

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How can I get better separation of my bands?

Product FAQ

Answer

First check the percentage of your agarose gel. A higher percentage will help you to resolve smaller molecular weights while a lower percentage will help you to resolve larger molecular weights.

Answer Id: E8010

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What are the TBE gel specifications, specifically the gel size and cassette size?

Product FAQ

Answer

Please see the gel specifications below:

Gel matrix: Acrylamide/bis-acrylamide
Gel thickness: 1.0 mm and 1.5 mm
Gel size: 8 cm x 8 cm (height x width)
Cassette size: 10 cm x 10 cm (height x width)

Answer Id: E7994

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My band is not moving through the nucleic acid gel during electrophoresis. What should I do?

Product FAQ

Answer

Check the power supply to ensure current is being passed through the gel. The running buffer composition must match the gel chemistry.

Answer Id: E8011

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What is the sensitivity of GelCode Color Silver Stain?

Product FAQ

Answer

Although detection will vary from protein to protein, this product will detect down to 0.1 ng of most proteins. The thicker the gel, the greater the sensitivity will be.

Answer Id: E8347

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What should the running conditions be for the TBE gels (voltage, current, run time, etc.)?

Product FAQ

Answer

Voltage: 200 V constant*
Approximate current at start: 10-18 mA/gel
Approximate current at end: 4-6 mA/gel
Run time: Approximately 30-90 minutes, dependent on gel percentage. The run is complete when the bromophenol blue (darker) tracking dye reaches the bottom of the gel.

* Voltages up to 250 V may be used to reduce run time.

Answer Id: E7995

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My bands are fuzzy when performing nucleic acid electrophoresis. What could cause this?

Product FAQ

Answer

Usually, fuzzy bands are caused by incorrect buffer solutions. Check the pH and salt concentrations in the running and gel buffers. Fuzzy bands can also be caused by excess salt in the samples or by old gels.

Answer Id: E8012

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What is the shelf life of GelCode Color Silver Stain Kit?

Product FAQ

Answer

The GelCode Color Silver Stain Kit is stable at room temperature (RT) for one year. All working solutions, except for the stabilizer, must be made immediately before each use. The stabilizer can be prepared in advance and stored at RT for several months.

Answer Id: E8348

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Can I stain my TBE gel or my TBE-urea gel? How?

Product FAQ

Answer

Yes, for ethidium bromide staining, soak the gel in a 2 μg/mL solution of ethidium bromide in ultrapure water for 20 minutes. Destain by rinsing with three successive 10-minute rinses of ultrapure water. Visualize bands under UV light

Answer Id: E7996

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I accidently allowed my DNA to run past the collection wells for my E-Gel™ SizeSelect™ or E-Gel™ CloneWell™ gels. What should I do?

Product FAQ

Answer

You can use the Reverse E-Gel™ Program to run the band back into the collection gel.

Answer Id: E8030

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My bands are too close together when performing nucleic acid electrophoresis. What can I do about this?

Product FAQ

Answer

We would recommend running the gel for longer or choosing a different gel that will have higher resolution in the size range you are targeting.

Answer Id: E8013

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Can the bacluovirus infect mammalian cells? How about Drosophila cells?

Product FAQ

Answer

Yes, baculovirus can infect mammalian cells, although only at very high titers. Baculovirus works best in liver cells. However, there is no danger of cross-contamination unless the cells are directly infected with the high-titer stocks. Bacuolvirus can infect Drosophila cells; however, it will not replicate in these cells. The promoters used to drive expression of your gene in a typical baculovirus system are both late promoters and require earlier proteins from the baculovirus genome. Thus, they will not work in S2 cells since the early proteins are not made.

Answer Id: E9433

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