Invitrogen BW1000 User manual

For Research Use Only. Not for use in diagnostic procedures.
Bandmate™Automated Western Blot
Processor
USER GUIDE
For automation of western blot processing
Catalog Numbers BW1000
Publication Number MAN0018722
Revision C.0

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The information in this guide is subject to change without notice.
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LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR
ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Revision history: Pub. No. MAN0018722
Revision Date Description
C.0 14 Sep 2021 Added to products for separate purchase and troubleshooting topics.
Minor corrections.
B.0 06 Oct 2020 Corrected safety and regulatory topics per guidance from Compliance.
Added Canadian French translations to Electrical Safety topic.
A.0 28 Feb 2020 New Manual.
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©2021 Thermo Fisher Scientific Inc. All rights reserved.

Contents
■CHAPTER 1 Bandmate™Automated Western Processor ....................... 4
Product description ............................................................. 4
Contents ....................................................................... 5
Operate the Bandmate™Automated Western Blot Processor ......................... 6
Set up the system .......................................................... 6
Program the processor ...................................................... 7
Run the processor .......................................................... 9
Clean the processor ........................................................ 10
General maintenance ....................................................... 10
Instrument dimensions and specifications .................................... 12
Troubleshooting ................................................................ 12
■APPENDIX A Safety ............................................................... 15
Symbols on this instrument ..................................................... 15
Standard safety symbols ................................................... 15
Control and connection symbols ............................................. 16
Conformity symbols ........................................................ 16
Safety information for instruments not manufactured by Thermo Fisher Scientific ...... 17
Instrument safety .............................................................. 17
General ................................................................... 17
Physical injury ............................................................. 18
Electrical safety ........................................................... 18
Cleaning and decontamination .............................................. 19
Instrument component and accessory disposal ................................ 19
Safety and electromagnetic compatibility (EMC) standards .......................... 19
Safety standards .......................................................... 19
EMC standards ............................................................ 20
Environmental design standards ............................................. 21
Chemical safety ................................................................ 22
Biological hazard safety ......................................................... 23
■APPENDIX B Documentation and support ...................................... 24
Customer and technical support ................................................. 24
Limited product warranty ........................................................ 24
Bandmate™Automated Western Blot Processor User Guide 3

Bandmate™Automated Western
Processor
Product description
The Invitrogen™Bandmate™Automated Western Blot Processor is a self-contained western blot
processing system that can process up to 4 mini blots or 2 midi blots at a time. Each blot can be
probed with dierent antibodies, if desired. The instrument has built-in traditional programs, but permits
custom programs for incubation and wash times. Antibodies and reagents are directly streamed into
incubation trays, which eliminates cross-contamination between reagents. The curved mini and midi
trays allow for even processing of blots with minimal antibody volumes; minimally 3.5 mL primary
antibody for mini blots and 7.5 mL for midi blots.
1
6
2
3
4
5
7
8
Figure 1 Instrument overview
1Reagent tubes (antibody reservoirs)
2Wash buer tubing
3Rocker cradle
4Tray cover
5Sample tray
6Reagent slider
7Waste and recovery funnel tray
8Waste tank compartment (behind door panel)
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4Bandmate™Automated Western Blot Processor User Guide

Table 1 Program icons
Icon Function Icon Function
Accept Fill
Back Home
Cancel (Stop) Pause
Continue Run
Drain Save
Edit
Contents
Table 2 Bandmate™Automated Western Blot Processor (Cat. No. BW1000)
Contents Amount
Buer Hose w/ Quick-connect Coupling 1
Interior Siphon Hose for Buer Bottle 1
Reagent Tubes 40
50 mL Recovery Tubes 8
Mini Trays 2
Midi Trays 2
Tray Covers 2
Funnel Trays 2
Waste Trays 2
Quick Reference Guide 1
Table 3 Available for separate purchase
Product Cat. No.
Reagent Tubes (100 tubes) BW020X5
Reagent Tubes (500 tubes) BW020X25
Mini Trays (2 trays) BW0060
Midi Trays (2 trays) BW0070
Chapter 1 Bandmate™Automated Western Processor
Contents 1
Bandmate™Automated Western Blot Processor User Guide 5

Table 3 Available for separate purchase (continued)
Product Cat. No.
Waste Tray BW0030
Funnel Tray BW0040
Tray Cover BW0050
4 L Buer Bottle BW0010
Table 4 Accessory products
Product Cat. No.
Pierce™Clear Milk Blocking Buer (10X) 37587
Blocker FL Fluorescent Blocking Buer (10X) 37565
Blocker BSA (10X) in PBS 37525
Blocker BSA (10X) in TBS 37520
SuperSignal™West Pico PLUS Chemiluminescent Substrate 34579
SuperSignal™West Atto Ultimate Sensitivity Substrate A38555
Pierce™Reversible Protein Stain Kit for Nitrocellulose
Membranes
24580
Pierce™Reversible Protein Stain Kit for PVDF Membranes 24585
Operate the Bandmate™Automated Western Blot
Processor
Set up the system
1. Set up the processor on a stable, level lab bench.
2. Open the door and remove the funnel tray.
3. Replace the funnel tray so that it hooks onto the catch at the back of the unit. Close the door.
4. Attach the wash buer hose to the connector located in the back of the instrument and attach the
other end to the buer bottle using the quick-connect fitting.
5. Plug the power supply into the power jack on the back of the instrument and connect the plug into
a wall outlet.
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Operate the Bandmate™Automated Western Blot Processor
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6Bandmate™Automated Western Blot Processor User Guide

Program the processor
•Store up to 8 programs.
•All incubation steps can be set from 1 second to 96 hours.
•Volume range for washes is between 10 mL to 75 mL for mini gels and 10 mL to 150 mL for midi
gels.
•We recommend using 20-30 mL per wash for mini gels and 30-50 mL for midi gels.
•Recommended antibody volumes are ≥3.5 mL for mini blots and ≥7.5 mL for midi blots.
•Maximum number of given steps in a protocol is 9.
•Recommended rocks per minute for incubation steps is 15 rpm. Recommended rocks per minute
for wash steps is 20 rpm.
1. From the home screen, select Programs. A list of programs, with default times will appear. The
Standard Western Blotting Protocol may be used for guidance and as a template. Select the
desired program using the dial. Press Edit.
2. Enter a name for the custom protocol. Select letters and numbers by turning the dial. Use the
cluster of 4 buttons at the lower left of the screen to navigate between characters and delete
characters or add spaces. When finished, press Save to save the program name, and then
press Continue .
3. Select whether the protocol will be used with a mini tray (two chambers with a central divider) or a
midi tray (one large chamber). Press Save , then press Continue .
4. Choose whether a quick rinse step is required in the protocol. A rinse will dispense 10 mL of wash
buer at the end of each step. If a rinse is desired, highlight Enable on the screen. Press Continue
.
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Bandmate™Automated Western Blot Processor User Guide 7

5. On the last screen, input the times and reagent for each step in your protocol. Buer wash steps
can be repeated up to ten times, but reagent steps cannot be repeated. When selecting the step,
the following may be chosen: Buer; Reagent 1; Reagent 2; and End. Use the toggle button to
highlight what needs to be modified and use the knob to move through the selections. Selecting
End indicates that there are no more steps. End must be selected as the last step in the protocol.
Below are the Preprogrammed Standard Western Blot Protocols, whereby Step 1 already has the
blocking solution pre-filled in the tray and contains two reagent (antibody addition) steps. Step 2
is the dispensing step for primary antibody, which is designated as Reagent 1, while Step 4 is the
dispensing step for secondary antibody and is designated as Reagent 2. Steps 3 and 5 are wash
steps with buer.
Table 5 Standard Mini
Step Reagent Repeat Fill volume Rock time Rocks per
min. Drain to[1]
1Pre Fill 1X — 1 hr 15 Waste
21 1X — 1 hr 15 Waste
3Buer 3X 20 mL 10 mins 20 Waste
42 1X — 30 mins 15 Waste
5Buer 6X 20 mL 5 mins 20 —
[1] Change from Waste to Recover if antibodies need to be saved.
Table 6 Standard Midi
Step Reagent Repeat Fill volume Rock time Rocks per
min. Drain to[1]
1Pre Fill 1X — 1 hr 15 Waste
21 1X — 1 hr 15 Waste
3Buer 3X 40 mL 10 mins 20 Waste
42 1X — 30 mins 15 Waste
5Buer 6X 40 mL 5 mins 20 —
[1] Change from Waste to Recover if antibodies need to be saved.
6. Choose the antibody recovery option if antibodies need to be recovered for future use. Recovered
reagents will be diverted to 50-mL conical tubes that can be placed in the brackets on either side
of the waste tray in the numbered holes (position 1 for first antibody addition and position 2 for
second antibody addition). If antibodies do not need to be recovered, all reagent steps may be set
to Waste. Press Save .
Note: If you choose to recover the antibody, place uncapped 50-mL recovery tubes, or
comparable 50-mL conical tubes, in the appropriate location(s) in the waste tray. Skirted conical
tubes are recommended because these tubes maintain proper positioning under the funnel tray
opening. If non-skirted tubes are used, align tubes vertically and do not allow the tubes to lean.
This will reduce the risk of solution loss during the recovery steps.
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8Bandmate™Automated Western Blot Processor User Guide

Note: When modifying an existing program, it is important to save before starting the program.
Only saved in-process programs will automatically resume after a power shutdown or outage.
Run the processor
1. Turn the power switch located at the back of the unit to the "On" position. The display will
illuminate and the moving parts will return to their home positions. After a few seconds, the home
screen will display.
2. Program or select your preferred western blot protocol (see “Program the processor” on page 7).
3. Fill the buer bottle in the back of the processor. Ensure that the hose hanging from the cap
reaches the bottom of the bottle. Leave the cap loose to allow air into the bottle as liquid is
pumped out.
For each wash, 10–75 mL of wash buer can be dispensed for mini gels and 10–150 mL for midi
gels. We recommend using 20–30 mL per wash for mini gels and 30–50 mL for midi gels. Ensure
there is enough wash buer to complete the protocol.
4. Flush the buer lines by selecting Flush from the home screen menu, then press Accept . Press
Fill until wash buer is dispensed from all nozzles. This may take several seconds. The wash
buer dispenses into the blot tray. Drain all wash buer from blot tray before use.
5. If using primary and secondary antibodies, two reagent tubes will be required for each blot being
processed. Check to ensure the foil on the bottom of the reagent tubes is not damaged. Screw
one reagent tube firmly and completely into the primary antibody position, noted with the number
1 toward the back of the slider. Next, screw the other reagent tube into the secondary antibody
position, noted with the number 2, just behind the buer dispensing nozzle.
6. Pipet the appropriate antibody solution into each reagent tube.
A standard-sized mini blot processed in a mini tray requires a minimum of 3.5 mL of each primary
antibody and secondary antibody solutions. A standard-sized midi blot processed in a midi tray
requires a minimum of 7.5 mL of each primary antibody and secondary antibody solutions.
7. Place blot(s) into the sample tray protein side up with protein lanes perpendicular to the tray gates.
If using a mini tray, which has a central divider, make sure the blot is on the same side of the
divider as the appropriate antibody reagent tubes.
8. Pour blocking buer of choice in to the tray. Use no more than 45 mL of blocking buer per mini
tray and no more than 90 mL of blocking buer for midi trays. Replace the tray cover and make
sure it is fully seated.
Tray Size Minimum Volume Maximum Volume
Mini 3.5 mL 45 mL
Midi 7.5 mL 90 mL
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Bandmate™Automated Western Blot Processor User Guide 9

9. To recover the antibody solutions, place empty 50 mL uncapped conical tubes in the appropriate
spaces on each side of the waste tray (space #1 for the primary antibody and #2 for the secondary
antibodies).
Note: For antibody recovery from midi trays, conical tubes must be added on both sides of the
waste tank as recovered solutions will be distributed in both tubes.
10. Check the waste tank to make sure that it is empty. Ensure that the funnel tray and waste trays are
pushed all the way to the back and are in place. Close the waste compartment and the work area
window.
11. Select the desired protocol from the menu and press Run .
12. Select the trays to be used for the protocol. Follow the instructions on the screen to indicate which
section(s) are in use.
13. Press Continue .
14. Press Run .
15. Once the run is complete, press Home to stop the rocker and return to the home screen.
Open the window and remove the blot. The blot is now ready for downstream processing
(chemiluminescent or colorimetric detection) and/or immediate imaging (fluorescence).
Note: The rocker will continue to rock with the final dispensed wash buer volume in the tray after
all of the steps of the protocol are completed to ensure that the blots do not dry out. The Home
button should be selected before removing the blot from the blot tray.
Clean the processor
1. Wash the sample trays, funnel trays, and tray covers using a gentle laboratory detergent. Rinse
with distilled water and dry face down.
2. Remove the Reagent Tubes from the instrument.
3. If the processor is not going to be used again for some time, flush the buer line with distilled
water to prevent salt build-up. To flush, place the wash buer hose into a container of distilled
water, then select Flush to run water through the lines and into the trays. Once water has been
flushed through the tubing, remove the tubing from the water container, then select Flush until the
liquid is completely flushed from the system and tubing.
General maintenance
Replacing buers
When replacing or adding additional wash buer to the wash bottle, it is recommended to flush the lines
to ensure no bubbles were introduced when the buer was replenished.
Chapter 1 Bandmate™Automated Western Processor
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10 Bandmate™Automated Western Blot Processor User Guide