GenScript eZwest Lite User manual

eZwestTM Lite
Automated Western Device
Start Your Easy Western from Here

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Table of Contents
Components and Initial setup
Warranty
1. Instrument Overview
1.1 Instrument overview
1.2 Specifications
1.3 Ordering information
1.4 Safety & Maintenance
2. Instructions
2.1 Instrument display and feature location
2.2 Basic operation
Save cassette ID when using the cassette for the first time
View and modify methods
Set cassette specific methods
2.3 Using the Pre-programmed blotting method
3. Sample Protocols
3.1 Quick Protocol
3.2 Normal Protocol
3.3 Overnight Protocol
4. Troubleshooting and FAQs
4.1 Error messages
4.2 FAQ
5. Technical Support
Appendix
A-1. Buffer recipes
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Version 20220615

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eZwestTM Lite Automated
Western Device
Initial setup
eZwestTM Lite Automated Western Device
eZwest Membrane Cassette
eZwest Cleaning Cassette
Stylus Pen
Power Cord
Forceps
Roller
Silicone Tubing
Waste Bottle
Wash Buffer Bottle
50 mL Tube
Membrane Spacer Sample
Quick Guide
Manual
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Description
Check and make sure the power plug matches the outlet
Place eZwestTM Lite device on a steady and levelled lab bench
Keep the vicinity of the instrument clean and ventilated
Make sure the power switch is OFF before plug in the instrument
Locate the three ports on the back of the device and connect them to
corresponding bottles with supplied tubing: wash buffer port to wash buffer
bottle, waste 1 and waste 2 ports to waste bottle
Power on the system
The device should beep and start
self-check. Once the screen displays
the main menu as shown on the right,
the unit is ready to go.
Important! Please check that all parts listed below are included with your package.
Components and Initial setup
L00816
L00820
L00821
L00822
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1 unit
1 unit
1 unit
1 pc
1 pc
1 pc
1 pc
3 pcs
1 pc
1 pc
3 pc
10 pcs
1 pc
1 pc
Components Cat. Size Quantity
Note: Before relocating the instrument, please drain liquid in the 50 ml tubes.
For the best result, please follow
these instructions to set up the
device before your first experiment.

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Warranty
GenScript warrants that eZwest
TM
Lite Automated Western Device is free from defects
in material and workmanship for a period of one year from the date of purchase or an
accumulative working time of 10,000 minutes, whichever comes first. If any defects
occur during this warranty period GenScript will at its option, repair, replace, or refund
the purchase price of the product at no charge to you.
Note: Damage caused by improper transportation, or any of the following actions are excluded:
• Improper operation.
• Repair or modification done by any other party than GenScript or an authorized
agent.
• Use of fittings or other spare parts supplied by any other party than GenScript.
• Damages caused by disasters.
For consultation and maintenance services, please contact GenScript’s customer
service and provide the following information.
Instrument model:
___________________________
Instrument serial number:
___________________________
Order number:
___________________________
Date of purchase:
___________________________

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Features and Benefits Hands-free: Press “START” and walk away
Easy-to-use: Set up in less than 10 minutes
Reliable: Comparable results to conventional manual method
Flexible: Fully customized procedures and solutions
Antibody Saver: Recovery of primary antibody
Multi-Blot Processing: Incubate up to two membranes at once
1.2 Specifications eZwestTM Lite Automated Western Device
Weight：
Dimensions(LxWxH)：
Electrical Requirements：
Operating Temperature：
Max Membrane Size(WxH)：
Throughput：
Application：
eZwest Lite：
Forceps：
Roller：
6.48 Kg
230×230×230 mm, 9.06x9.06x9.06 in
100-120 V or 220-240 V，50/60 Hz
15-40°C, 59-104°F
90x83 mm, 3.5x3.2 in
1 channel for up to 2 membranes
Western blotting
ABS，PP，stainless steel，silicone
ABS
Stainless steel, POM
1. Instrument Overview
1.1 Instrument
Overview
eZwestTM Lite is a fully automated blotting device that frees researchers from the
manual labor of traditional western protocol. eZwestTM Lite can be programmed to
run a sequence of western blot membrane incubation steps automatically including:
blocking, primary antibody and secondary antibody incubations and washing. When
desired, the primary antibody can be recovered for re-use. The processed
membranes are ready for detection. Automation of the incubation steps minimizes
errors and variability typically associated with manual operations contributing to
experimental reproducibility.
The eZwestTM Lite device is compatible with the traditional reagents and protocols
used for conventional western blotting. No proprietary reagents are required. Fully
customizable steps and solutions make eZwestTM Lite a powerful tool fitting specific
experiment needs for both routine and novel western blotting conditions.

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1.4 Safety & Maintenance
To ensure optimal performance,
we recommend regularly
cleaning the eZwest instrument.
Component
eZwest device: routine
cleaning with the
membrane cassette
eZwest device:
extensive cleaning with
the cleaning cassette
Membrane cassette
Note
Description
After each use, please clean the device following the procedure
below.
1. Insert the membrane cassette into the device.
2. Go to “Method”
3. Select “System Clean”
4. Add 20 mL wash buffer (PBST or TBST) in the Blocking buffer
tube. Keep the 1° Ab and 2° Ab tubes clean and empty.
5. Ensure there is at least 100mL wash buffer in the wash buffer
bottle and the Waste bottle is empty. Press OK to confirm the
buffer volume message.
6. Start “System Clean” method by clicking OK again.
7. After the cleaning procedure is finished, dispose the waste
solution.
After every 20 uses or every 2 weeks，perform the extensive
cleaning following the procedure below.
1. Empty the Wash bottle and Waste bottle
2. Add 100 mL wash buffer in the Wash bottle
3. Insert the cleaning cassette into the device.
4. Go to “Method”
5. Select “System Clean”
6. Add 20 mL of eZwest cleaning solution (Appendix A-1) in the
Blocking buffer tube. Keep the 1° Ab and 2° Ab tubes clean and
empty. Click OK to confirm the buffer volume message.
7. Start “System Clean” program by clicking OK again.
8. After the cleaning procedure is finished, remove the cleaning
cassette and dispose the waste solution.
9. Perform the routine cleaning protocol once.
Open the membrane cassette and rinse with distilled water or 10%
ethanol/IPA and let it dry after each use.
If the device will be left unused for over 2 weeks,
please ensure the tubes in the device are filled with regular wash
buffer (1X PBST or TBST with 20 mM EDTA).
1.3 Ordering information
eZwest Membrane Cassette
(included in L00816)
Product
eZwest Membrane Cassette
Size
1 unit
Cat.
L00820
Accessories and other
reagents
Quantity
1
Product
eZwest Cleaning Cassette
Membrane Spacer
Stylus Pen
Size
1 unit
50 pcs
1 pc
Quantity
1
1
1
Cat.
L00821
L00824
L00822

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2. Instructions
2.1 Instrument display and feature location
eZwestTM Lite automated
western device Touch screen Stylus pen
Front cover Reagent chamber
AC power receptacle
System ON/OFF switch
Wash buffer
Waste 1
Waste 2
Blocking
buffer
1°Ab
2°Ab
Front view
Back view
Top view
Fig. 1
Fig. 2
Fig. 3

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Screen display (main menu) Current program name
System menu Method menu
Current program
steps
Program timer
Start program
Screen display (Method view) Exit
Device programs
System clean
Cassette program
Program steps
Select program Save program
Membrane cassette
Latch
Buffer ports
Sensor chip
Fig. 4
Fig. 5
Fig. 6

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2.2 Basic operation
Save cassette ID when using the
cassette for the first time
1. Switch on the device.
2. Insert the membrane cassette into the device as shown in Fig. 7. Make sure the pins
are pushed all the way in, and the side with the “eZwest” logo is upright.
Note: The pins are designed to be movable as part of the auto-seal mechanism. They may
appear to be in different position after removing the cassette, which is normal.
3. After the cassette is inserted, you will be asked if you want to “save ID”.
4. Press OK to save cassette ID as show in Fig. 8.
Note: Each eZwest Lite device could save up to 5 cassette IDs.
View and modify methods 1. Enter method setting by pressing "Method” on the upper right corner of the main menu.
Fig. 7
Fig. 8
Fig. 9

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2. Select method on the left to show the steps of each method.
3. Select a step to show and modify the associated details.
Note: Each Method may be programmed with 6 steps.
4. Each step within a Method is customizable :
a. Reagent option.
b. Number of Cycles required to repeat the step.
Fig. 10
Fig. 11
Fig.12
Fig. 13

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c. Run time for each cycle in minutes.
d. Pump Speed for recirculating the liquid flow through the membrane cassette.
5. Once the method is set up, press “Save”
Set cassette specific methods The eZwestTM Lite device can store up to one dedicated method for each cassette. Upon
cassette installation, select the associated method.
1. Method settings can be changed as described above
2. Change name of membrane cassette
a. Enter Method Setting and select the name of membrane cassette
b. Press “Edit ID” and change name with the pop-up keyboard.
Fig.14
Fig.15
Fig.16
Fig.17

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2.3 Using the Pre-programmed blotting method
1. Prepare blotting solutions
a. Blocking buffer, 10 mL
b. Primary antibody solution, 10 mL
c. Secondary antibody solution, 10 mL
d. Wash buffer, 10 mL per wash cycle, e.g. 90 mL if wash 3 times and each
with 3 cycles
2. Place the prepared solutions into eZwestTM Lite device as indicated below.
Note: Assure all fittings and caps are tightly sealed.
3. Open the membrane cassette: release the latch by rotating it up as shown in Fig. 19(i),
lift the latch and pull it towards the top of the cassette as in Fig. 19(ii), the opened
cassette is shown in Fig. 19(iii).
Blocking buffer
1°Ab
2°Ab
Wash buffer Waste2Waste1
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iii
Fig. 18
Fig.19

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7. Check and ensure all the tubes and bottles contain adequate amount of reagents.
Warning: insufficient buffer or incorrect solution placement may result in experiment failure.
8. Select the method on the main menu and click “Start” to run.
9. Once the procedure is finished, click “Finish”, and replace the membrane cassette with
the cleaning cassette. Membranes are ready for detection.
10.Replace all solutions in the reagent chamber with wash buffer or cleaning solution to
start the system cleaning cycle when prompted according to the cleaning guidance in
1.4 Safety and Maintenance.
4. Placing membrane(s) in the casette
a. Processing a single membrane:
Place the membrane in the center of the cassette surface as shown below in
Fig. 20(i), with the protein side facing up and the high MW side aligning towards
the bottom edge of the cassette as Fig. 20(ii). Place a spacer across the high
MW end (bottom) of the membrane along the membrane edge.
b. Processing two membranes simultaneously:
Place the first membrane as described in Fig. 20(i). Then place a second spacer
across the low MW side of the first membrane along the membrane edge. Lay
the second membrane on top with the protein side facing down and high MW
side align towards the bottom edge of the cassette. Make sure both spacers are
in between membranes.
Note: The spacers are reusable for up to 20 times. Replace spacer if deformed
or contaminated.
Membrane 2
2nd Spacer
Membrane 1
1st Spacer
1st Spacer
Membrane 1
Low M.W.
High M.W.
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ii
5. Carefully close the cassette and lock the latch.
6. Securely insert the membrane cassette into the device as shown below, make sure the
side with “eZwest” logo is facing up-right.
Fig. 20
Fig. 21

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3. Sample Protocols
3.1 Quick Protocol: Requires sensitive antibodies and/or higher abundant proteins. All steps performed at
room temperature.
Step
Blocking
Wash
1°Antibody
Wash
2°Antibody
Wash
Cycles
1
3
1
3
1
3
Time
30min
5min
60min
5min
30min
5min
Pump speed
Slow
Normal
Slow
Normal
Slow
Normal
3.2 Normal Protocol: All steps performed at room temperature.
Step
Blocking
Wash
1°Antibody
Wash
2°Antibody
Wash
Cycles
1
3
1
3
1
3
Time
60min
5min
120min
5min
60min
5min
Pump speed
Slow
Normal
Slow
Normal
Slow
Normal
3.3 Overnight Protocol:
Use the protocol for very low abundant proteins and/or antibodies with low affinity.
Note: Overnight protocol can ONLY be saved and run as a cassette method.
Step
Blocking
Wash
1°Antibody
Wash
2°Antibody
Wash
Cycles
1
3
Overnight
3
1
3
Time
60min
5min
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5min
60min
5min
Pump speed
Slow
Normal
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Normal
Slow
Normal
1. Press “Method” button on the main interface.
Fig. 22

Traget overnight step
Blocking
1°Ab
2°Ab
Type of reagent needed
Blocking, Wash
Blocking, Wash, 1°Ab
Blocking, Wash, 1°Ab, 2°Ab
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2. Select the cassette ID (e.g., “345”) to enter the cassette program.
3. Select the steps for customization (overnight option is only available to Blocking, 1°Ab,
and 2°Ab steps).
4. Open the dropdown menu for “Cycles” and select “Overnight”. The same setting can be
applied to 1°Ab or 2°Ab steps.
5. Press “Save” button to save program.
6. Press “Use” to use this method and back to main interface.
7. The initial required reagents for different overnight processes are shown in the table.
The loading volume for each reagent is the same as the normal protocol. Press “Start” to
initiate the process.
Fig. 23
Fig. 24
Fig. 25
Fig. 26
Fig. 27

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8. Check buffer volumes for each required reagent.
E.g., if select Overnight Cycle under the 1°Ab step, only Wash buffer, Blocking buffer
and 1°Ab solution are required to start the overnight protocol in the eZwest device.
9. After 1° Ab incubation step is finished in eZwest, the machine will automatically pause
with a notice as shown in Fig. 29.
10. Remove the membrane cassette from eZwest and put it overnight at 4°C.
11. A system cleaning cycle is required after the membrane cassette is removed. Insert a
cleaning cassette or any other empty cassette for the routine cleaning cycle.
12. The system will first recover the antibodies if Antibody Recovery is selected in the
method setting.
13. After recovery is complete, the antibody solution can be stored away for reuse.
Fig. 28
Fig. 29
Fig. 30
Fig. 31
Fig. 32

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14. Replace the blocking buffer with 20mL wash buffer and leave antibody tubes empty for
the cleaning cycle. Check the buffer volumes and start system cleaning.
15. Put the overnight cassette back into eZwest. The device will recognize the unfinished
overnight protocol and request to continue. Click “OK” to continue or “Cancel” to abort.
16. As the blocking and primary antibody incubation steps are completed before the
overnight process, prepare only wash buffer and secondary antibody solution for the
remaining process. If antibody recovery is selected, put an empty tube for the
recovered primary antibody. After checking the volumes by clicking “OK”, the program
will resume once “Start” is pressed.
Fig. 33
Fig. 34
Fig. 35
Fig. 36

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4. Troubleshooting and FAQs
4.1 Error messages Error Message
RFID Error
No Cassette
Pump Error
System Error
Possible causes
Can’t read cassette chip/no
cassette
Cassette not secured in
position
Pump malfunction
System malfunction
Solutions
Replace cassette/ insert
cassette
Check Pins and Reinsert
cassette securely
Contact GenScript
Contact GenScript
4.2 FAQ Description
Strong background
Weak signal
Fluorescent detects only
strong signal around
spacer
Possible causes
Insufficient wash
1. Antibody is too diluted
2. Instrument needs
cleaning
3. Insufficient antibody
incubation time
Weak target signal
contrasted out by
interference
Solutions
1. Increase number of wash
cycles (set to 5 cycles for 3
minutes each)
2. Increase pump flow per wash
cycles (set to normal)
1. Reduce antibody dilution factor
2. Clean eZwestTM Lite instrument
3. Increase antibody incubation
time or reduce incubation
pump flow
1. Adjust detection area to omit
spacer position
2. Cover spacer signal region
with paper or tape

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5. Technical Support
Visit the GenScript web site at www.genscript.com for:
1. Technical resources, including manuals, vector maps and sequences, application notes,
MSDSs, FAQs, formulations, citations, handbooks, etc.
2. Complete technical support contact information
3. Access to the GenScript Online Catalog
4. Additional product information and special offers
For more information or technical assistance, call, write, fax, or email.
GenScript USA Inc.
860 Centennial Ave. Piscataway, NJ 08854
Tel: [phone]
FAX: [phone]
Email: [email protected]