Invitrogen™ E-PAGE™ Midi Protein Gels, 3.7 mm

Catalog No.	Wells	Gel Percentage
EP09606	96-well	6%
EP04808	48-well	8%

Catalog No. EP09606 Supplier Invitrogen™ Supplier No. EP09606

Description

Includes

8 E-PAGE™ 96 gels, 4.5mL E-PAGE™ Loading Buffer 1 (4X), 1 Butterfly Opener, 1 E-PAGE™ Blotting Pad

E-PAGE™ Gels are self-contained, pre-cast gels that include a gel matrix and electrodes packaged inside a disposable cassette.

E-PAGE Gels are self-contained, pre-cast gels that include a gel matrix and electrodes packaged inside a disposable cassette. Protein separation on an E-PAGE Gel requires an EBase Integrated Device, a compact and automated platform designed for medium- and high-throughput electrophoresis.

Each E-PAGE 8% 48-well gel contains 48 sample lanes and 4 marker lanes with a 3.2-cm separation length. E-PAGE 48-well gels can be loaded with a 12- or 8-channel pipettor or 4- or 8-tip robotic system (Figure 1).

Each E-PAGE 6% 96-well gel contains 96 sample lanes and 8 marker lanes with a 1.6-cm separation length. The well openings of the cassette are compatible with the standard 96-well plate format and can be conveniently loaded with a multichannel pipettor or 8-, 12-, or 96-tip liquid-handling robotic device (Figure 2).

Both E-PAGE 48 and 96 cassettes can be opened to remove the gel for downstream staining or blotting (Figures 3 and 4). In addition, each cassette is labeled with an individual barcode to facilitate identification of the gel using commercial barcode readers.

More about the E-PAGE precast gel system
The Invitrogen E-PAGE precast gel system is designed for fast, mess-free medium- and high-throughput protein electrophoresis. It consists of the E-PAGE precast gels, E-Base Integrated devices for running the gels, and free E-Editor 2.0 software for image analysis.

The all-in-one E-Gel Power Snap Plus Electrophoresis System is designed for fast and convenient high-throughput gel electrophoresis. With dry precast E-PAGE gel technology, you can run protein samples in as little as 14 minutes. The device arrives with pre-programmed protocols for both E-PAGE gel types.

Order Info

Shipping Condition: Room Temperature

Specifications

• Gel Percentage: 6%
• Gel Size: Midi
• Gel Thickness: 3.7 mm
• Gel Type: E-PAGE
• Sample Loading Volume: Up to 20 μL
• Separation Range: 10 to 300 kDa
• Separation Type: Denaturing
• Wells: 96-well
• Mode of Separation: Molecular Weight
• Quantity: 8 gels (1 kit)
• Shipping Condition: Room Temperature
• Storage Requirements: Store at room temperature, do not allow the temperature to drop below 4°C or rise above 40°C
• Length (Metric): 8 cm
• Width (Metric): 13 cm
• Product Line: E-PAGE
• For Use With (Equipment): E-BASE

Frequently Asked Questions (FAQs)

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

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).

What is the difference between the EG and EP program for the E-Base system?

The EG program is to run E-Gel 96 and 48 gels, while the EP is to run the E-PAGE 96 and 48 gels.

What sample loading buffer should be used for E-PAGE gels?

For SDS-PAGE and staining or blotting, we recommend using the 4X E-PAGE Loading Buffer 1 for preparing samples. We do not recommend using any other SDS-PAGE sample buffer. The E-PAGE Loading Buffer 1 is optimized for E-PAGE gels. If the sample is already in NuPAGE LDS sample buffer or the Tris-Glycine SDS sample buffer, it is fine to run on the E-PAGE gels. However, there may be a loss of resolution in the bands.

Where can I download the latest version of the E-Gel 96 analysis software?

The E-Editor 2.0 Software can be downloaded for free from the Thermo Fisher Scientific website (search for 'e-editor 2.0').

Are the E-PAGE Gels (example Cat. No. EP09606) compatible with the iBlot3 device (Cat. O. IB31001)?

Due to the thickness of the E-PAGE gels, they are not compatible with the iBlot3 device for transfer. Recommended methods for transfer of E-PAGE gels are semi-dry blotting or semi-wet blotting in the XCell II Blot Module. Methods are described in the E-PAGE Technical Guide.

I transferred my proteins from an E-PAGE gel using the iBlot Dry Blotting System and my protein bands are distorted on the membrane. Why is this?

This indicates that a non-uniform electric field was created around the wells. Ensure that the well protrusions on the E-PAGE gel are properly flattened using the De-bubbling Roller. To ensure the best blotting results, we recommend using the De-bubbling Roller with E-PAGE gels. If you use the Blotting Roller with E-PAGE gels, be sure to follow the recommendations on page 22 of the manual (https://tools.thermofisher.com/content/sfs/manuals/iblotsystem_man.pdf) to obtain good results.

What transfer conditions do you recommend for semi-dry transfer of E-PAGE gels using the Invitrogen Semi-Dry Blotter?

For semi-dry transfer of E-PAGE 48 gels, we recommend using 2X NuPAGE Transfer Buffer containing 15% methanol and for E-PAGE 96 gels, we recommend using 2X NuPAGE Transfer Buffer without methanol. Transfer at 25 V (constant) for 30-60 minutes. Please refer to Page 33 of the E-PAGE Technical Guide (http://tools.thermofisher.com/content/sfs/manuals/epagetechguide_man.pdf).

Can E-PAGE gels be transferred using a semi-dry blotting protocol?

The preferred method of transfer for E-PAGE gels is dry blotting using the iBlot/iBlot 2 Dry Blotting System (see Page 27 of the E-PAGE Technical Guide http://tools.thermofisher.com/content/sfs/manuals/epagetechguide_man.pdf). Please note that this works only for E-PAGE 48 gels. Semi-dry blotting using the Invitrogen Semi-Dry Blotter, Cat. No. SD1000 (Page 33), or semi-wet blotting (Page 37) may also be performed.

Can I use the iBlot Filter Paper for blotting E-PAGE gels?

We do not recommend using the iBlot Filter Paper for blotting E-PAGE gels.

What method do you recommend for blotting E-PAGE gels?

The preferred method of transfer for E-PAGE gels is dry blotting using the iBlot/iBlot 2 Dry Blotting System (see Page 27 of the E-PAGE Technical Guide http://tools.thermofisher.com/content/sfs/manuals/epagetechguide_man.pdf). Please note that this works only for E-PAGE 48 gels. Semi-dry blotting using the Invitrogen Semi-Dry Blotter, Cat. No. SD1000 (Page 33), or semi-wet blotting (Page 37) may also be performed.

What kind of regular maintenance do you recommend for the Mother and Daughter E-Base devices?

We recommend keeping the surfaces of the Mother E-Base Device and Daughter E-Base Device free of contaminants. To clean, disconnect bases from power source and wipe with a dry cloth. Do not attempt to open or service the bases.

What is the purpose of the E-Holder Platform?

The E-Holder Platform is designed to hold E-PAGE Gels during loading. You can use the E-Holder Platform when you need to load multiple gels while other gels are running on the E-Base device.

Note: The E-Holder Platform is not a power supply unit, cannot be connected to an electrical outlet, and cannot be used to run E-PAGE 48 or 96 Gels. To obtain the best results, run E-PAGE Gels on the Mother E-Base Device or Daughter E-Base Device within 15 minutes after loading on the E-Holder Platform.

What is the recommended program to be used for running E-PAGE gels on an E-Base device?

We recommend using the EP program for running E-PAGE gels.

How do you recommend staining E-PAGE gels?

E-PAGE gels are compatible with many standard Coomassie staining, silver staining, or fluorescent staining protocols. E-PAGE gels are thicker than most SDS-PAGE mini-gels, so additional time may be required for staining and destaining steps. We recommend the following stains for E-PAGE Gels. Detailed staining protocols are described on Pages 40-49 of the E-PAGE Technical guide (http://tools.thermofisher.com/content/sfs/manuals/epagetechguide_man.pdf).

Total protein stains:
*SYPRO Ruby Protein Gel Stain (Page 40)
*Coomassie Stain (Page 42)
*SimplyBlue SafeStain (Page 44)
*SilverQuest Silver Staining Kit (Page 47)
*SilverXpress Silver Staining Kit (Page 48)

Specific protein stains:
*Lumio Green Detection Reagent for detecting Lumio fusion proteins Ppage 40)
*InVision His-tag In-Gel Stain for detecting 6X His-tagged proteins (blotting recommended first, Page 49)

How can I open my E-PAGE gels after electrophoresis?

After electrophoresis is completed, the E-PAGE gel cassette can be opened with a gel knife to perform downstream applications such as western blotting or staining.

Which protein standards do you recommend using with E-PAGE 48 and E-PAGE 96 gels for electrophoresis, Western blotting and fluorescent detection?

Electrophoresis:
For E-PAGE 48 8% gels, use SeeBlue Plus 2 Pre-Stained Standard, Cat. No. LC5925.
For E-PAGE 96 6% gels, use E-PAGE SeeBlue Pre-Stained Standard, Cat. No. LC5700.

Western Blotting:
For E-PAGE 48 8% gels, use MagicMark XP Western Protein Standard, Cat. No. LC5602.
For E-PAGE 96 6% gels, use E-PAGE MagicMark Unstained Protein Standard, Cat. No. LC5701.

Fluorescence detection:
For E-PAGE 48 8% gels, use BenchMark Fluorescent Protein Standard, Cat. No. LC5928.
For E-PAGE 96 6% gels, use BenchMark Fluorescent Protein Standard, Cat. No. LC5928.

What is the recommended run time for E-PAGE 96 6% and E-PAGE 48 8% gels?

The recommended run time for E-PAGE 96 6% gels is 14 minutes and for E-PAGE 48 8% gels is 25 minutes. No pre-run is necessary. It is possible to run longer than the recommended run time to improve resolution, keeping in mind the risk of running into the next well below. Avoid running E-PAGE 96 or 48 gels for more than 25 and 30 minutes respectively. Running power is 9.5 watts.

What do you recommend for drying my stained E-PAGE gel?

We recommend using the Large Gel Drying Kit (Cat. No. NI2207) for drying E-PAGE gels. It is also possible to air dry E-PAGE gels. The E-PAGE 96 gels will need at least 4 days for complete air drying. Vacuum drying is not recommended - the thickness of these gels makes them especially prone to cracking.

How much salt or detergent can I have in my samples to be loaded on your E-PAGE gels?

Please follow the guidelines mentioned on Page 10 of the E-PAGE Technical Guide (http://tools.thermofisher.com/content/sfs/manuals/epagetechguide_man.pdf). Samples containing high salt or detergents result in loss of resolution on E-PAGE Gels.

What is the composition of the 4X E-PAGE Loading Buffer 1?

The composition of the 4X E-PAGE Loading Buffer 1 is proprietary

I have Lumio labeled proteins and would like to run them on E-PAGE 48/96 gels. Which sample buffer should I use?

We recommend using the Lumio gel sample buffer that is supplied with the Lumio Green Detection kit, for in-gel detection. We do not recommend using the E-PAGE loading buffer. There is no need to remove the E-PAGE gels from the cassette to visualize Lumio fusion proteins.

Which sample buffer should I use for preparing my samples to be run on your E-PAGE gel?

We recommend using the 4X E-PAGE Loading Buffer 1 (supplied with E-PAGE gels) for preparing samples for SDS-PAGE and staining or blotting using E-PAGE gels. We do not recommend using any other SDS sample buffer such as the NuPAGE LDS sample buffer or the Tris-Glycine SDS sample buffer as there may be a loss of resolution in the bands.

What is the recommended total sample loading volume per lane of an E-PAGE gel?

The recommended total sample loading volume for E-PAGE 48 and E-PAGE 96 gels is 15 µL (up to 20 µL). Very low volumes of sample loaded will result in poor resolution and smearing. For proper band separation, we recommend keeping sample volumes uniform and loading deionized water into the empty wells.

What is the recommended protein amount to be loaded per lane of an E-PAGE gel and what is the maximum recommended protein amount per lane?

We recommend loading 1-20 ?g protein per lane of an E-PAGE gel. The maximum recommended protein load per lane gel is 20 µg. Excess protein will cause poor resolution and smearing.

Do E-PAGE gels contain SDS?

E-PAGE gels contain SDS and are recommended for performing electrophoresis under denaturing conditions.

What is the running distance for E-PAGE 96 6% and E-PAGE 48 8% gels?

The running distance of each lane is 1.6 cm for E-PAGE 96 6% and 3.2 cm for E-PAGE 48 8% gels.

What is the molecular weight range of proteins that can be separated on E-PAGE gels?

The molecular weight range of proteins that can be separated is 10-220 kDa for E-PAGE 96 6% gels and 10-200 kDa for E-PAGE 48 8% gels.

Do E-PAGE gels contain a preservative?

E-PAGE gels do not contain a preservative. They undergo UV sterilization during the production process.

What is the formulation of E-PAGE gels?

The gel formulation of E-PAGE gels is proprietary.

What is special about E-PAGE gels?

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.