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Manual / Product Insert

Novex Pre-Cast Gel Electrophoresis Guide

Version: MAN0003187 Rev date: 29 October 2010
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Product FAQ

What gels can I use to separate native proteins?

Answer

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.

Answer Id: E5195

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Product FAQ

What are the recommended sample loading volumes and protein loading amounts for IEF gels?

Answer

The recommended sample loading volumes and protein loading amounts for the different well formats are listed on Page 8 of the Invitrogen Pre-Cast Gel Electrophoresis Guide (https://tools.thermofisher.com/content/sfs/manuals/electrophoresisguide_man.pdf).

Answer Id: E10633

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Product Literature

NuPAGE® Novex® Pre-Cast Gels - Green Fact Sheet

Product FAQ

What are the recommended sample loading volumes and protein loading amounts for Tricine gels?

Answer

The recommended sample loading volumes and protein loading amounts for the different well formats are listed on Page 8 of the Invitrogen Pre-Cast Gel Electrophoresis Guide (https://tools.thermofisher.com/content/sfs/manuals/electrophoresisguide_man.pdf).

Answer Id: E10721

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Product FAQ

What are the recommended sample loading volumes and protein loading amounts for Zymogram gels?

Answer

The recommended sample loading volumes and protein loading amounts for the different well formats are listed on Page 8 of the Invitrogen Pre-Cast Gel Electrophoresis Guide (https://tools.thermofisher.com/content/sfs/manuals/electrophoresisguide_man.pdf).

Answer Id: E10728

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Product FAQ

My PowerEase 90W Power Supply stopped functioning with an alarm and the screen displays “No Load”. What is possibly wrong?

Answer

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.

Answer Id: E11018

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Product FAQ

My ZOOM Dual Power Supply stopped running with an alarm and the screen display shows “NO LOAD”. What is the problem?

Answer

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.

Answer Id: E11052

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Product FAQ

What is the chemistry of your Invitrogen Tris-Glycine gels?

Answer

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.

Answer Id: E10555

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Product FAQ

My PowerEase 300W Power Supply stopped functioning with an alarm and the screen displays “No Load”. What is possibly wrong?

Answer

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.

Answer Id: E11008

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Product FAQ

What are the applications that your IEF gels can be used for?

Answer

*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.

Answer Id: E10628

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Product FAQ

Is it possible to run a denaturing IEF gel?

Answer

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.

Answer Id: E10645

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