Documents & Support

To help you choose the TBE polyacrylamide gel that will provide optimal separation results for your nucleic acid fragment, we have designed gel migration charts for each gel type offered. Please check the TBE gel migration charts on page 74 of the Novex Pre-Cast Gel Electrophoresis Guide (https://tools.thermofisher.com/content/sfs/manuals/electrophoresisguide_man.pdf).

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

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Here are possible causes and solutions:

- High voltage application is set to run on a very low current. DISABLE No Load alarm on the Running Screen (see manual for details), for example when performing an isoelectric focusing application.
- Electrophoresis leads are not connected to the power supply or to the electrophoresis unit(s), or there is a broken circuit in the electrophoresis cell. Check the connections to the power supply and on your electrophoresis cell to make sure the connection is intact; check condition of wires in electrophoresis unit. Close the circuit by reconnecting the cables. Press Stop/Start to restart the run.
- High resistance due to tape left on a pre-cast gel, incorrect buffer concentration, or incorrect buffer volumes in the electrophoresis cell. Correct the condition by making sure the tape is removed from the pre-cast gel, buffers are prepared correctly, and the recommended volume of buffer is sadded to the electrophoresis unit.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Here are the possible causes and suggested solutions:

- Electrophoresis leads are not connected to the power supply or to the electrophoresis unit(s), or there is a broken circuit in the electrophoresis cell. Check the connections to the power supply and on your electrophoresis cell to make sure the connection is intact; check condition of wires in electrophoresis unit. Close the circuit by reconnecting the cables. Press Start/Pause to restart the run.
- High resistance due to tape left on a pre-cast gel, incorrect buffer concentration, or incorrect buffer volumes in the electrophoresis cell. Correct the condition by making sure the tape is removed from the pre-cast gel, buffers are prepared correctly, and the recommended volume of buffer is added to the electrophoresis unit.
- Current has dropped below acceptable rating (4 mA). The power supply has a built-in “No Load Detection” feature that can detect when the current drops below the acceptable rating of 4 mA.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Here are possible causes and solutions:

- Electrophoresis leads are not connected to the power supply or to the electrophoresis unit(s), or there is a broken circuit in the electrophoresis cell. Check the connections to the power supply and on your electrophoresis cell to make sure the connection is intact; check condition of wires in electrophoresis unit. Close the circuit by reconnecting the cables. Press Start/Pause to restart the run.
- High resistance due to tape left on a pre-cast gel, incorrect buffer concentration, or incorrect buffer volumes in the electrophoresis cell. Correct the condition by making sure the tape is removed from the pre-cast gel, buffers are prepared correctly, and the recommended volume of buffer is added to the electrophoresis unit.
- Current has dropped below acceptable rating (10 mA). The power supply has a built-in “No Load Detection” feature that can detect when the current drops below the acceptable rating of 10 mA.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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The NativePAGE Invitrogen Bis-Tris Gel System is a pre-cast polyacrylamide mini gel system that provides a sensitive and high-resolution method for analyzing native membrane protein complexes, native soluble proteins, molecular mass estimations, and assessing purity of native proteins. It is based on the blue native polyacrylamide gel electrophoresis technique (BN PAGE) developed by Schagger and von Jagow.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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Invitrogen Tris-Glycine polyacrylamide gel chemistry is based on the Laemmli system with minor modifications for maximum performance in the pre-cast format. These gels do not contain SDS and can therefore be used to accurately separate both native and denatured proteins by using the appropriate running buffer. Invitrogen Tris-Glycine Gels provide reproducible separation of a wide range of proteins into well resolved bands.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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*Invitrogen Pre-Cast Vertical IEF gels are excellent for native applications using soluble proteins.
*They can be used for pI determination and confirmation of isoforms of purified products.
*They can readily detect minor changes in a protein due to deamination, phosphorylation or glycosylation, and can resolve different proteins of similar size, which cannot be resolved on standard SDS-PAGE gels. Pre-focusing is not required.
*For a quick 2D analysis, proteins can be separated by pI in the first dimension, using IEF gels, and then by mass in the second dimension, using NuPAGE Bis-Tris gels or Invitrogen Tris-Glycine gels with a 2D-well or by using ZOOM gels for 2D SDS-PAGE.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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After electrophoresis, the gel should be stained in a 0.5-1.0 µg/mL solution of ethidium bromide in deionized water for 15 to 60 min depending on the thickness of the gel. As an optional step to reduce background fluorescence, the gel can be destained in deionized water for 15 to 30 min. Alternatively, ethidium bromide may be added directly to the agarose prior to casting. Add ethidium bromide to melted agarose to a final concentration of 0.5 µg/mL. This has the advantage of reducing the amount of ethidium bromide waste. However, this procedure may reduce the migration rate of nucleic acids.

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It is possible run a denaturing IEF gel, but our Invitrogen pre-cast vertical IEF gels were not designed for denaturing applications. You can try adding ~7 M urea to the sample prior to running to help solubility. This may work for some proteins but not for others. Further, the protein may re-precipitate since there is no urea in the gel itself, resulting in fuzzy bands and smearing. Use of urea in IEF gels or in running buffers is not generally common because the urea does not remain dispersed throughout the gel. It will migrate to a certain point in the gel and then stop moving.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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E-PAGE gels are self-contained, pre-cast gels (neutral pH) that include a buffered gel matrix and electrodes packaged inside a disposable, UV-transparent cassette. They are designed for fast, buffer-less, high-throughput protein electrophoresis in a horizontal format and are available in 96-well, 6% and 48-well, 8% formats. Each E-PAGE 96 6% gel contains 96 sample lanes and 8 marker lanes in a patented staggered well-format that is compatible with the standard 96-well plate format for loading with a multichannel pipette or with an automated liquid handling system. Each E-PAGE 48 8% gel contains 48 sample wells and 4 marker wells. The wells are compatible for loading with a multichannel pipette in alternating lanes or with an automated liquid handling system.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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We do offer disposable, empty plastic cassettes and combs to provide the option of pouring your own Mini or Midi gels for use with the XCell SureLock Mini-Cell, Mini Gel Tank, or XCell4 SureLock Midi-Cell, respectively. All cassettes are sealed on three sides to prevent leaking. Since the cassettes are pre-sealed with the slot pre-taped, a bulky casting stand is not necessary. The empty cassettes and combs can be found here (https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-gel-electrophoresis/power-supplies-protein-electrophoresis.html#staining).

- The Empty Mini Gel Cassettes are available in two thicknesses (1.0 mm and 1.5 mm) and can be used with Invitrogen Combs that are available in two thicknesses (1.0 mm and 1.5 mm), and in 7 types (1 well, 2 well, 5 well, 10 well, 12 well, 15 well, and 2D well).
- The Bolt Empty Mini Gel Cassettes are available in 1.0 mm thickness and can be used with Bolt 10- or 12-well combs.
- The Empty Midi Gel Cassettes are available in 1.0 mm thickness and can be used with three Invitrogen Comb types (12 + 2 wells, 20 wells, and 26 wells).

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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The E-PAGE SeeBlue Prestained Standard is designed for use with E-PAGE pre-cast gels, to visualize protein molecular weight ranges during electrophoresis and evaluate western transfer efficiency. The apparent molecular weights of the proteins in the E-PAGE SeeBlue Prestained Standard are shown below:

- Myosin: 261 kDa in 6% E-PAGE gel
- Phosphorylase B: 173 kDa in 6% E-PAGE gel
- BSA: 97 kDa in 6% E-PAGE gel
- Alcohol dehydrogenase: 42 kDa in 6% E-PAGE gel
- Myoglobin: 21 kDa in 6% E-PAGE gel

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

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The Thermo Scientific Power Blotter (Cat. No. PB0012) consists of the Thermo Scientific Power Station (Cat. No. PB0010) with activated Blotting Software and the Thermo Scientific Power Blot Cassette (Cat. No. PB0002). It is designed for rapid semi-dry transfer of proteins from polyacrylamide gels to nitrocellulose or PVDF membranes. Traditional western blotting techniques require a transfer of one hour to overnight to achieve good transfer efficiency. When used in conjunction with Thermo Scientific 1-Step Transfer Buffer, the Thermo Scientific Power Blotter is designed to provide transfer efficiency in 5-10 min that is equivalent to, or better than, traditional blotting techniques without the need for gel pre-equilibration. This significant reduction in protein transfer time is accomplished by optimizing the ionic strength of the transfer buffer and increasing the current [amps (A)/cm2] flowing through the transfer stack. The system has been verified to work with commonly used pre-cast and homemade SDS-PAGE gels. The Thermo Scientific Power Blotter can also be used for standard semi-dry transfer protocols with Towbin buffer.

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

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