ThermoFisher Scientific E-PAGE Gels Quick start guide

E-PAGE™Technical Guide
General information and protocols for using E-PAGE™Gels
Catalog Number EP096-06
Revision B.0
Publication Number MAN0000374
Publication Part Number 25-0644
For Research Use Only. Not for use in diagnostic procedures.

For support visit www.lifetechnologies.com/support or email [email protected]om
www.lifetechnologies.com
06 October 2021

iii
Contents
Product Contents........................................................................................................................................ iv
Introduction .................................................................................................................................1
E-PAGE™Protein Electrophoresis System ................................................................................................1
E-PAGE™System Components ..................................................................................................................3
Methods .......................................................................................................................................8
Electrophoresis of E-PAGE™Gels.........................................................................................8
Guidelines for Sample Preparation............................................................................................................8
Electrophoresis of E-PAGE™Gels ............................................................................................................13
Running E-PAGE™Gels.............................................................................................................................17
Guidelines for Loading E-PAGE™Gels by Automated Liquid Handling ..........................................18
Using E-Holder™Platform ........................................................................................................................20
Opening the E-PAGE™Cassette ...............................................................................................................24
Blotting E-PAGE™Gels.........................................................................................................25
Blotting E-PAGE™Gels with the iBlot®Device ......................................................................................25
Semi-Dry Blotting of E-PAGE™Gels........................................................................................................31
Semi-Wet Blotting of E-PAGE™Gels .......................................................................................................35
Visualizing and Staining of E-PAGE™Gels ........................................................................38
Visualizing LumioFusion Proteins .......................................................................................................39
SYPRO®Ruby Staining Protocol ..............................................................................................................40
Coomassie Staining ....................................................................................................................................41
SimplyBlue™ SafeStain ...............................................................................................................................43
Silver Staining Methods.............................................................................................................................45
InVision™His-tag Protein Gel Stain.........................................................................................................48
Drying of E-PAGE™Gels ......................................................................................................49
Gel Drying ...................................................................................................................................................49
Analysis and Troubleshooting ............................................................................................51
Molecular Weight Calibration ..................................................................................................................51
Expected Results.........................................................................................................................................53
Using E-Editor™2.02 Software..................................................................................................................60
Troubleshooting..........................................................................................................................................61
Appendix....................................................................................................................................63
Additional Staining Protocols...................................................................................................................63
Product Specifications................................................................................................................................67
Accessory Products ....................................................................................................................................70
Technical Support.......................................................................................................................................72

iv
Product Contents
Purpose of the
Guide
The E-PAGE™Technical Guide contains information about E-PAGE™Gels and
is intended to supplement the manuals supplied with E-PAGE™Gels. Details
for sample preparation, electrophoresis conditions, blotting, and staining are
included in this guide.
Types of Kits
E-PAGE™gels are designed for use with the E-Base™ Electrophoresis Device,
and are available in 48-well or 96-well formats. See pages 3–4 for further
specifications. See page 71 for ordering information.
Product Quantity Catalog no.
E-PAGE™48 8% Gels 8-pack EP048-08
E-PAGE™96 6% Gels 8-pack EP096-06
Kit Contents
Each pack of E-PAGE
™
Gels contain the following contents:
E-PAGE™Gel 8-Pack Contents Quantity
E-PAGEGels 8 gels
E-PAGELoading Buffer 1 (4X) 4.5 mL
E-PAGEBlotting Pad 1
Shipping and
Storage
The E-PAGE™48 and 96 Gels, E-PAGEBlotting Pad and Mother E-Base™are
shipped and stored at room temperature.
Do not allow the temperature to drop below 4°C or rise above 40°C when
storing the gels.
The E-PAGELoading Buffer is shipped at room temperature. Store the
E-PAGE™Loading Buffer 1 (4X) at room temperature or at 4°C.

1
Introduction
E-PAGE™Protein Electrophoresis System
About the
E-PAGE™Protein
Electrophoresis
System
The E-PAGE™Protein Electrophoresis System is designed for fast, medium-to-
high-throughput protein electrophoresis in a horizontal format.
The self-contained E-PAGE™48 or 96 Pre-cast Gels consist of a buffered gel
matrix and electrodes packaged inside a disposable, UV-transparent cassette.
The gels can be loaded by multichannel pipetter or automated loading system.
Electrophoresis of the samples is performed with the E-Base™Electrophoresis
Device. The E-Base™Electrophoresis Device consists of a base for
electrophoresis, and an integral power supply. After electrophoresis is
completed, the cassette can be opened with the Invitrogen™Gel Knife (Cat.
No. EI9010, sold separately) to perform downstream applications such as
western blotting or staining.
The E-Editor™2.02 software allows digital images of the E-PAGE™Gels to be
reconfigured into a side-by-side format for easy comparison and analysis.
Applications
The E-PAGE™Protein Electrophoresis System is ideal for screening protein
samples using the following applications:
•In-gel staining with Lumio™Green Reagent
•Western blotting
•Staining (Coomassie, silver, or fluorescent stains)
•Functional assays
Advantages
Using E-PAGE™Gels for protein analysis offers the following advantages:
•Produces fast, safe, and consistent electrophoresis
•Eliminates the need to prepare gels and buffers
•Compatible with most commercially available automated liquid handling
systems for high-throughput electrophoresis
•Available in two well formats to suit your applications
•Easy to open gel cassettes facilitate downstream staining and blotting
applications
•Ability to track samples during electrophoresis of multiple gels using an
individual barcode on each gel cassette

2
E-PAGE™Protein Electrophoresis System, Continued
Choosing a Gel
for your
Application
To obtain the best results for your application it is important to choose the
correct E-PAGE™Gel. The table below describes the major features of each
format. For specifications and separation range of each gel type, see page 67–68.
E-PAGE™ Gel No. Rows No. Wells Well Volume Run
Length
Run
Time
48 Gel 2* 48 sample
4 marker
15 µL
15 µL 3.2 cm 25 min.
96 Gel 8* 96 sample
8 marker
20 µL
20 µL 1.6 cm 14 min.
*Wells compatible for loading with a multichannel pipetter or with an automated
liquid handling system.
Western Blotting
of E-PAGE™Gels
E-PAGE
™
Gels are suitable for western blotting. The iBlot
®
Gel Transfer Device
is recommended method for blotting E-PAGE™Gels (page 25). Semi-dry
blotting (page 25), and semi-wet blotting may also be performed (page 35).
Tank blotting is not recommended, but can be performed if even pressure is
maintained between the gel and the membrane, and cooling is provided.
Staining E-PAGE™
Gels
E-PAGE
™
Gels are compatible with many standard Coomassie, silver, 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 38–48.
Total Protein Stains:
•SYPRO® Ruby Protein Gel Stain (page 38)
•Coomassie Stain (page 41)
•SimplyBlue™SafeStain (page 43)
•SilverQuest™ Silver Staining Kit (page 46)
•SilverXpress® Silver Staining Kit (page 47)
Specific Protein Stains:
•Lumio™Green Detection Reagent for detecting Lumiofusion proteins
(page 38)
•InVision™His-tag In-Gel Stain for detecting 6X His-tagged proteins
(blotting recommended first, page 48)

3
E-PAGE™System Components
Introduction
The E-PAGE™Protein Electrophoresis System consists of the following
components:
•E-PAGE™48 or 96 Pre-cast Gels
•E-Base™Electrophoresis Device
•E-Holder™Platform
•E-PAGE™Loading Buffer 1 (4X)
•E-PAGE™ Blotting Pad
•E-Editor™2.02 Software
Details on each system component are described in this section.
E-PAGE48 Gels
•
E-PAGE™48 Gels are self-contained, pre-cast gels that include a buffered
gel matrix and electrodes packaged inside a disposable, UV-transparent
cassette.
•Each E-PAGE™48 Gel contains 48 sample lanes and 4 marker lanes. This
configuration provides a 3.2-cm run length. The wells of E-PAGE™48 Gels
are compatible for loading with a multichannel pipetter in alternating lanes
or with an automated liquid handling system (see page 17 for automation
specifications).
•After electrophoresis, the E-PAGE™48 cassette is easily opened with the
Invitrogen™Gel Knife (Cat. No. EI9010, sold separately) to remove the gel
for staining or blotting applications.
•Each E-PAGE™48 cassette is labeled with an individual barcode to facilitate
identification of the gel using commercial barcode readers (page 13).
See page 67 for E-PAGE™48 8% Gel specifications.
Diagram of an
E-PAGE

48
Cassette
A diagram of the E-PAGE
™
48 Gel cassette is shown below.
Plastic combs
Copper
electrode (-)
Copper
electrode (+)
Barcode

4
E-PAGE™System Components, Continued
E-PAGE96 Gels
•E-PAGE
™
96 Gels are self-contained, pre-cast gels that include a buffered
gel matrix and electrodes packaged inside a disposable, UV-transparent
cassette.
•Each E-PAGE™96 Gel contains 96 sample lanes and 8 marker lanes in a
patented staggered well format that provides a 1.6-cm run length. The wells
of the E-PAGE™96 gels are compatible for loading with a multichannel
pipetter or with an automated liquid handling system (See page 17 for
automation specifications).
•After electrophoresis, the E-PAGE™cassette is easily opened with the
Invitrogen™Gel Knife (Cat. No. EI9010, sold separately) to remove the gel
from the cassette for staining or blotting applications.
•Each E-PAGE™96 cassette is labeled with an individual barcode to facilitate
identification of the gel using commercial barcode readers (page 13).
See page 67 for E-PAGE™96 6% Gel specifications
Diagram of an
E-PAGE™96
Cassette
A diagram of the E-PAGE™96 Gel cassette is shown below.
Copper
electrode (+)
Copper
electrode (-)
Barcode
Plastic combs

5
E-PAGE™System Components, Continued
E-Base
Electrophoresis
Device
E-PAGE™Gels are used with a specially designed electrophoresis device that
combines a base and a power supply. Two types of devices are available from
Life Technologies:
•The Mother E-Base™(Cat. no. EB-M03) has an electrical plug that can be
connected directly to an electrical outlet and is used for electrophoresis of
one E-PAGE™Gel. The Mother E-Base™has been tested for electrophoresis
with up to three Daughter E-Bases™connected at one time.
•The Daughter E-Base™(Cat. no. EB-D03) connects to the Mother E-Base™,
and together they can be used for the independent electrophoresis of 2 or
more E-PAGE™Gels. The Daughter E-Base™does not have an electrical plug
and cannot be used without a Mother E-Base™.
E-PAGE
™
Gels are not compatible with the E-Gel
®
96 bases
(Cat nos. G7100-01/G7200-01) previously available from Life Technologies. The
older E-Gel®96 bases do not have the ‘E-Base™’ inscription on the platform.
Mother E-Base™
Each Mother E-Base
™
has a pwr/prg (power/program) button (right side) and a
time button (left side) on the lower-right side of the base. The lower-left side of
each Mother E-Base™contains a light LED and a digital display. The gel cassette
is inserted into the two electrode connections. The Mother E-Base™is connected
to an electrical outlet with the electrical plug.
The E-Base™EP program is set to 14 minutes default time. See page 15 to adjust
time as recommended.
Mother E-Base

(–)
(+)
Pwr/prg button
Time button
Digital
display
To electrical outlet
LED Light

6
E-PAGE™System Components, Continued
Daughter E-Base™
The Daughter E-Base™is similar to the Mother E-Base™ except the Daughter
E-Base™ does not have an electrical cord and cannot be connected to an electrical
outlet.
The Daughter E-Base™ is connected to a Mother E-Base™or to another Daughter
E-Base™(already connected to a Mother E-Base™). Once connected to a Mother
E-Base™, each Daughter E-Base™ is designed to function independently of the
Mother E-Base™ or other Daughter E-Bases™
Mother E-Base™/Daughter E-Base™
Daughter
E
-Base™
Mother
E
-Base™