Invitrogen Bandmate MAN0018722, User Manual

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5.
On the last screen, input the times and reagent for each step in your protocol. Buffer wash steps
can be repeated up to ten times, but reagent steps cannot be repeated. When selecting the step,
the following may be chosen:
Buffer ; 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 buffer.
Table 5 Standard Mini
Step Reagent Repeat Fill volume Rock time
Rocks per
min.
Drain to
[1]
1 Pre Fill 1X — 1 hr 15 Waste
2 1 1X — 1 hr 15 Waste
3 Buffer 3X 20 mL 10 mins 20 Waste
4 2 1X — 30 mins 15 Waste
5 Buffer 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]
1
Pre Fill 1X — 1 hr 15 Waste
2 1 1X — 1 hr 15 Waste
3 Buffer 3X 40 mL 10 mins 20 Waste
4 2 1X — 30 mins 15 Waste
5 Buffer 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.
Chapter 1
Bandmate
™
Automated Western Processor
Operate the Bandmate ™ Automated Western Blot Processor
1
8
Bandmate ™ Automated Western Blot Processor User Guide