GenScript eStain 2.0 User manual

Aug 15, 2014
GenScript USA Inc.
860 Centennial Ave.
Piscataway, NJ 08854
Tel: [phone]
Fax: [phone]
Email: [email protected]
For fast Coomassie blue staining of proteins in mini polyacrylamide gels
2.0
®

Table of Contents
Product Contents
Product Specifications
eStain®2.0 Protein Staining Device
Introduction
Quick Reference Guide
Protocols
Troubleshooting
Examples of Results
Technical Support
Warranty
Appendix
1
3
5
7
10
11
20
22
23
24
25
eStain®2.0 Protein Staining System www.genscript.com

Product Contents Product Contents, continued
Type of Products
eStain®2.0 Protein Staining
Device Contents
Upon Receiving
the Instrument
eStain®Protein Staining
Pads
eStain®Protein Staining
Pads, continued
Product
eStain®2.0 Protein Staining Device
Component
eStain®2.0 Protein Staining Device
eStain®2.0 Graphite Electrode
(Installed inside the device )
Sponge Cushion
(Installed inside the device )
Regional Specific Power Cord
Forceps
Shovel
Tray
Quantity
1 each
1 each
2 each
1 each
1 each
1 each
1 each
Cat. No.
L02016
Product
eStain®Protein Staining Pads (R-250, 20-pak)
eStain®Protein Staining Pads (G-250, 20-pak)
Cat. No.
L02011
L02012
This manual is supplied with the following product:
The contents of the eStain®2.0 Protein Staining Device are listed below:
Examine the unit carefully for any damage incurred during transit. File any
damage claims with the carrier. The warranty does not cover in-transit
damage.
The following eStain®Protein Staining Pads are available from GenScript:
If you order the eStain®Protein Staining Pads, you will receive the components
listed in the table below. Store the eStain®Protein Staining Pads at room
temperature. For best results, use the eStain®Protein Staining Pads before the
expiration date printed on the package.
eStain®2.0 Graphite
Electrode Product
eStain®2.0 Graphite Electrode
Cat. No.
L02017
The following eStain®2.0 Graphite Electrode is available from GenScript:
For best results, after 100 uses of electric staining, replace the worn eStain®
2.0 Graphite Electrode with a new one.
Component
The eStain®Protein Staining Pads
(R-250, 20-pak) contains:
eStain®Protein Staining Pad (R-250)
Absorbent Filter Paper
Sponge Cushion
The eStain®Protein Staining Pads
(G-250, 20-pak) contains:
eStain®Protein Staining Pad (G-250)
Absorbent Filter Paper
Sponge Cushion
each pack of eStain®Protein Staining Pad
(R-250) contains:
1×eStain®Cathode Pad (R-250)
1×eStain®Anode Pad
each pack of eStain®Protein Staining Pad
(G-250) contains:
1×eStain®Cathode Pad (G-250)
1×eStain®Anode Pad
eStain®2.0 Protein Staining System www.genscript.com eStain®2.0 Protein Staining System www.genscript.com
Quantity
20
1
2
20
1
2
1
1
1
1
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Product Specifications Product Specifications, continued
eStain®2.0 Protein Staining
Device Specifications
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Weight:
Dimensions:
Electrical Parameters:
Built-in Features:
Compatibility:
Materials:
Operating Temperature:
Forceps:
Shovel:
Tray:
1.78 kg
325 mm (l) × 195 mm (w) × 70 mm (h)
100-120 V, 220-240 V, 50/60 Hz, 3.2 A
Digital display, alarm, light LED
Suitable for fast Coomassie blue staining of proteins in
mini polyacrylamide gels
Acrylonitrile butadiene styrene, Polycarbonate,
Aluminum, Titanium, Plasticized silicone.
Room temperature
Stainless steel
Polycarbonate
Polycarbonate
eStain®Protein Staining
Pads Specifications
size:
Materials:
90 mm (l) × 80 mm (w) × 2.5 mm (t)
Blotting filter paper presoaked with proprietary
cathode buffer containing CBB dye R-250 or G-250
size:
Materials:
90 mm (l) × 80 mm (w) × 2.5 mm (t)
Blotting filter paper presoaked with proprietary
anode buffer
The eStain®Protein Staining Pads are used with the eStain®2.0 Protein Staining
Device.
The specifications of the eStain®Protein Staining Pads are listed below:
eStain®Cathode Pad (R-250 or G-250)
Avoid acetone, dimethyl sulfoxide, and acetic acid. These reagents can erode or
damage the device.
eStain®Anode Pad
Size:
Materials:
80 mm (l) × 70 mm (w) × 2.6 mm (t)
Vegetable fiber
Absorbent Filter Paper
eStain®2.0 Graphite
Electrode Specifications
Dimensions:
Weight:
Materials:
100 mm (l) × 92 mm (w) × 8.8 mm (t)
206 g
Powdered carbon, Clay, Stainless steel
The eStain®2.0 Graphite Electrode is used as the replaceable anode
electrode of eStain®2.0 Protein Staining Device and available separately
from GenScript. The specifications for eStain®2.0 Graphite Electrode are
listed below:
eStain®Protein Staining
Pads Specifications,
continued
Size:
Materials:
104 mm (l) × 100 mm (w) × 5 mm (t)
Sponge
Sponge Cushion
eStain®2.0 Protein Staining System www.genscript.com eStain®2.0 Protein Staining System www.genscript.com

Control Panel
Open Button
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eStain®2.0 Protein Staining Device eStain®2.0 Protein Staining Device, continued
Front View of
eStain®2.0 Device
Rear View of
eStain® 2.0 Device
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Control Panel of
eStain®2.0 Device
Top View of
Openned eStain®2.0
Device
The control panel of the eStain®2.0 Protein Staining Device is described below.
The Digital Display shows two rows of multi-digits that specify the electric
staining conditions as follows:
The upper three digits after text <PN> indicate number of usage of the
eStain®2.0 Graphite Electrode.
The lower four digits specify the time of electric staining in minute and second.
The two status lights show the working modes of the eStain®2.0 Protein
Staining Device. When the right status light is on, the device is switched on
and working at staining mode; when both left and right status lights are on,
the device is working at numbering mode.
The Reset button is used to clear parameters.
The Min. button is used to shift between
staining and numbering mode, and to
set running time. Each short press will
increase one minute. Each long press
(2 seconds) will toggle working mode from
staining to numbering or vice versa.
The Sec. button is also used to set running
time, each press will increase 5 seconds.
The Start/Stop button is used to activate/stop
the staining program.
The top view of openned eStain®2.0 Protein Staining Device identifying
various parts is shown below.
The rear view showing various parts of the eStain®2.0 Protein Staining Device
is shown below.
The front top view showing various parts of the eStain®2.0 Protein Staining Device
is shown below.
eStain®2.0 Protein Staining System www.genscript.com eStain®2.0 Protein Staining System www.genscript.com
Power SwitchAC Inlet
Titanium Cathode
Graphite Anode

The eStain®2.0 Protein Staining System is based on GenScript’s
patent-pending electric staining technology.To use the eStain®2.0 Protein
Staining System for protein gel staining, assemble the eStain®Protein
Staining Pad with your pre-run gel on the eStain®2.0 Protein Staining Device.
Similar to semi-dry blotting, the eStain®Cathode Pad and eStain®Anode
Pad act as ion reservoirs with proprietary anode and cathode buffers. eStain®
Cathode Pad, presoaked with cathode buffer containing CBB dye R-250 or
G250, supplies negatively charged staining reagents. The voltage generated
between graphite anode and titanium cathode allows for rapid and directional
movement of negatively charged Coomassie blue dye into the gel matrix to
stain proteins as well as for the homogeneous movement of the unbound
staining reagents out of the gel matrix to destain the gel within only 7 minutes
or less. Therefore the eStain®2.0 Protein Staining System provides fast,
convenient and reliable in-gel protein staining without the need to prepare
additional buffers.
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eStain®2.0 Protein Staining System www.genscript.com
Introduction
System Overview
System Components The eStain®2.0 Protein Staining System consists of:
eStain®2.0 Protein Staining Device
The eStain®2.0 Protein Staining Device is a user-friendly electric staining unit that allows
for fast, convenient and efficient in-gel protein staining with Coomassie blue dye reagents.
eStain®Protein Staining Pads
The disposable eStain®Protein Staining Pads are used to perform electric staining of
proteins in gels with Coomassie blue dye. Two types of eStain®Protein Staining Pads,
R-250 and G-250, are available separately from GenScript and for different requirements.
Each pack of eStain®Protein Staining Pad contains an eStain®Cathode Pad presoaked
with proprietary cathode buffer containing CBB dye R-250 or G-250, and an eStain®
Anode Pad presoaked with proprietary anode buffer, allowing for rapid, convenient and
reliable in-gel protein staining without the need to prepare additional buffers.
The eStain®2.0 Protein Staining System, applying GenScript's patent-pending electric
staining technique, allows you to quickly, reliably and efficiently stain proteins in various
types of mini polyacrylamide gels with Coomassie blue dye. It consists of the eStain®2.0
Protein Staining Device and the eStain®Protein Staining Pads.The proprietary electric
staining technology of the eStain®2.0 Protein Staining Device combined with the eStain®
Protein Staining Pads applies a voltage generated between graphite anode and titanium
cathode to allow for quick and directional movement of negatively charged Coomassie
blue dye into the gel matrix to stain the proteins and also the homogeneous movement of
the unbound staining reagents out of the gel matrix to destain the gel matrix within only 7
minutes or less. The eStain®2.0 Protein Staining System integrates three steps of
conventional Coomassie blue staining method into a single one and greatly cuts down the
time required for protein staining analysis. The eStain®2.0 Protein Staining System is able
to deliver high detection sensitivity down to several ng per protein band, which is
comparable to the sensitivity of traditional method despite of their different working
principles.
Introduction, continued
System Mechanism

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eStain®2.0 Protein Staining System www.genscript.com
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eStain®2.0 Protein Staining System www.genscript.com
Introduction, continued
System Mechanism,
continued
System Features Important features of the eStain®2.0 Protein Staining System are listed below:
· Innovative electric staining technology created for fast, reliable and
efficient protein gel staining in 7 minutes or less.
· User-friendly electric staining unit allowing for easy and convenient
procedures.
· Consumable staining pads offering convenience without the need for
additional buffers.
· High staining efficiency as conventional Coomassie blue staining method.
· Compatible for use with various types of mini polyacrylamide gels.
Schematic mechanism of eStain®2.0 Protein Staining System showing the
movement of Coomassie blue dye:
eStain®Cathode Pad
eStain®Anode Pad
Pre-run gel
E
EEE
CBB
dye CBB
dye
CBB
dye
CBB
dye
CBB
dye
CBB
dye
Titanium Cathode
2e-+ 2H2O H2+ 2OH-
Graphite Anode
4OH-O2+ 2H2O + 4e-
Quick Reference Guide
Mode
Sponge cushion and
graphite electrode
installed inside
eStain®2.0 Device
eStain®2.0 Device
plugged in
eStain®2.0 Device
and staining stack
assembled
Time selection
Run
End of run
Checking number of
usage of graphite
anode
Replacement of worn
graphite anode
Action
Insert the sponge
cushion and graphite
electrode in the
anode tank
Connect eStain® 2.0
Device to an
electrical outlet and
power switch is on
Place staining stack
on the device and
close lid
Press Min. and Sec.
button to select
desired running time
Press Start/Stop
button
Automatic
Press and hold Min.
button for 2 seconds
Switch off the device
and replace the worn
graphite anode with a
new one
Sound
–
–
–
–
–
Continuous beeping
for 2 minutes
–
–
Light
–
Steady right
light
Steady right
light
Steady right
light
flashing right
light
Steady right
light
Steady left
and right
lights
–
Display
–
Default running
time (00:00)
Default running
time (00:00)
Running time
defined
Count down
time
Default running
time (00:00)
Number of usage
of the graphite
electrode
–
A quick reference guide for operating the eStain®2.0 Protein Staining Device
is provided below.
Introduction

1. Check the Power Cord supplied with the unit to ensure that the cord is
compatible with local socket format.
2. Place the eStain®2.0 Protein Staining Device on a leveled laboratory bench.
Keep the area around the device clear to ensure proper ventilation of the
unit.
3. For your safety: Position the device properly such that the power switch
and the AC inlet located on the rear of the unit are easily accessible.
4. Ensure that the AC power switch is in the Off position.
5. Open the closed lid of the eStain®2.0 Protein Staining Device by pressing
the Open button. Place one or two pieces of Sponge Cushion in the anode
tank depending on gel thickness. For 1.5 mm gel, use one piece of Sponge
Cushion; for 0.75 and 1.0 mm gel, use two pieces of Sponge Cushion.
6. Insert the eStain®2.0 Graphite Electrode into the anode tank as described
in Section "Replacing the eStain®2.0 Graphite Electrode", then close the
lid of the device.
eStain®2.0 Protein Staining System
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eStain®2.0 Protein Staining System www.genscript.com
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www.genscript.com
Protocols
Recommendations
Installing the
eStain®2.0 Device
Follow these recommendations for best results:
1. Wear gloves at all times during the entire staining procedures to prevent
contamination of pads and gels.
2. Avoid using expired eStain®Protein Staining Pads. Always use the pads
before the specified expiration date printed on the package.
Protocols, continued
Installing the
eStain®2.0 Device,
continued
Note: For a new eStain®2.0 Protein Staining Device, two pieces of Sponge Cushion and
one unit of eStain®2.0 Graphite Electrode have already been installed inside the
unit for immediate use. If staining 0.75 and 1.0mm gel, use one or two pieces of
Sponge Cushion; If staining 1.5mm gel, use one or zero piece of Sponge Cushion.
If use the gel of Invitrogen, must remove the bump at the bottom of the gel.
9. When the electrophoresis of your samples is almost completed, press
the power switch (located on the rear of the device) to turn ON the
eStain®2.0 Protein Staining Device. The right status light is on indicating
staining mode. The lower four digits of the digital display show the default
running time (00:00).
You are ready to use the eStain®2.0 Protein Staining Device for staining
applications.
7. Pull out the waste tray from the right side of the device. Place a new
Absorbent Filter Paper inside the tray and then push the tray back in.
Note: After 20 times of electric staining, replace the used Absorbent Filter Paper with
a new one. One piece of new Absorbent Filter Paper is included in each box of
eStain®Protein Staining Pads.
8. Attach the power cord to the AC inlet, then, to the electrical outlet.
Use only properly grounded AC outlets and power cords.
Note: After 20 times of electric staining, replace the used Sponge Cushions with new ones.
A pair of new Sponge Cushions are included in each box of eStain®Protein Staining
Pads.

Assembling Staining
Stack with eStain®
Pads and Pre-run Gel
1. Open the closed lid by pressing the Open button.
2. Remove one package labeled as eStain®Protein Staining Pad from the
eStain®Protein Staining Pads box and tear the laminated sealing of the
package. Remove the two small packages labeled 1×eStain®Cathode
Pad R-250 (or G-250), and 1×eStain®Anode Pad, respectively.
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eStain®2.0 Protein Staining System www.genscript.com
Protocols, continued
3. Tear the sealing of the 1×eStain®Anode Pad package. Remove the
eStain®Anode Pad from the package and place it on the anode plate of
the eStain® 2.0 Protein Staining Device.
5. Place the gel on the eStain®Anode Pad. Gently remove air bubbles
between gel and anode pad using the small shovel supplied with the
device.
4. Carefully remove the pre-run gel containing your protein samples from the
gel cassette and briefly rinse the gel with distilled water.
Protocols, continued
Assembling Staining
Stack with eStain®
Pads and Pre-run Gel,
continued
7. Press the Open button, then push back and close the lid of eStain®2.0
Protein Staining Device.
6. Tear the sealing of the 1×eStain®Cathode Pad package. Remove the
eStain®Cathode Pad from the package and place it on top of the gel.

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eStain®2.0 Protein Staining System www.genscript.com
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eStain®2.0 Protein Staining System www.genscript.com
Protocols, continued
Gel Thickness
0.75 mm
1.0 mm
1.5 mm
Running Time
6 - 7 minutes
7 - 8 minutes
7 - 8 minutes
Performing Electric
Staining
2. Press the Start/Stop button to activate the electric staining program. The running
time begins to count down and right status light keeps flashing during the whole
staining program.
After assembling the staining stack, perform electric staining as described below.
Perform electric staining within 15 minutes after assembling the pads with the gel.
1. Use the following parameters as starting points for electric staining based on gel
thickness.
Note: For best staining results, it is recommended to use eStain®2.0 Protein Staining System at
room temperature (22 - 28 ℃). If the ambient temperature is below 22 ℃, you may need to
extend the running time to obtain satisfied staining results based on gel thickness.
Press the Min. and Sec. buttons to set appropriate running time. If an
undesired running time is set by mistake, press Reset button to clear the
wrong time, and then press again the Min. and Sec. buttons to choose the
desired running time.
3. At the end of the staining, current automatically shuts off and the eStain®
2.0 Protein Staining Device signals the end of staining with repeated
beeping sounds. The right status light stops flashing and the lower four
digits show text (00:00).
4. Press any button on the control panel to stop the beeping.
5. Proceed to disassemble the stack and clean the device.
To obtain consistent staining results, disassemble the staining stack right
away after ending the staining procedure.
1. Open the closed lid by pressing the Open button.
2. Carefully separate the stained gel from the staining stack and proceed
to document the gel image.
3. Discard the used eStain®Protein Staining Pad.
Note: Do Not reuse the eStain®Protein Staining Pad. Discard after each use.
4. Take the eStain® 2.0Graphite Electrode out of the device, dry the contact
rods with a dry cloth or tissue paper. Take care not to lose the spring on
the rod located near the triangle symbols before placing back into the
device.
Caution: Do Not clean the eStain®2.0 Graphite Electrode immediately after ending the
staining program since the graphite electrode is still hot after a run. Wait for 3
to 5 minutes before proceeding to cleaning.
5. Clean the titanium cathode and the anode tank with a dry cloth or tissue
paper.
6. Replace the used Sponge Cushions with new ones after 20 times of
electric staining.
7. Replace the used Absorbent Filter Paper in the waste tray with a new
one after 20 times of electric staining.
At this point, the eStain®2.0 Protein Staining Device is ready for another run.
If you are not using the device for a period of time, turn off the power switch
located on the back of the device, take the graphite anode out, and keep the
lid open to let any solution left in the holes for the contact rods dry out.
For any other repairs and services, contact Technical Support. Do not perform
any repairs or services on the eStain®2.0 Protein Staining Device by yourself
to avoid damaging the device.
Disassembling and
Cleaning the
eStain®2.0 Device
Protocols, continued
Performing Electric
Staining, continued