Life technologies Invitrogen iBlot Reference guide

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REFERENCE
GUIDE
iBlot® Dry Blotting System
Catalog numbers: IB1001, IB1001EU, IB1001UK
Rev. Date: 23 February 2011
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REFERENCE
CARD
Part number 25-0912 MAN0003281
Intended Use: For research use only. Not intended for any animal or human therapeutic or diagnostic use.
For technical support, email [e-mail].
For country-specific contact information, visit www.invitrogen.com.
Instructions for using the iBlot® Gel Transfer Device to perform dry blotting of proteins from mini- or midi-gels with iBlot® Gel
Transfer Stacks is described below. For detailed instructions, including instructions for transfer of proteins from E-PAGE™ gels,
transfer of DNA from agarose gels or polyacrylamide gels to iBlot® DNA Transfer Stacks, and performing western detection with
iBlot® Western Detection Stack, refer to the manual supplied with the product or download the manual from www.invitrogen.com.
General Guidelines
• Remove air bubbles as indicated in the protocol using the Blotting Roller.
• Do not trim the membrane or iBlot® Gel Transfer Stacks to t your gel size. Use the iBlot® Gel Transfer Stacks, Regular for
blotting E-PAGE™, 1 midi-, or 2 mini-gels. Use iBlot® Gel Transfer Stacks, Mini for blotting 1 mini-gel.
• Pretreatment of the gel after electrophoresis is not required. Remove the gel after electrophoresis and proceed with blotting.
• It may be necessary to use a Run Time of 8–10 minutes for the transfer if your protein of interest is >150 kDa.
• You may need to reduce the Run Time to 5–6 minutes for the transfer if your protein of interest is <30 kDa.
1. Open the lid of the device. 2. Unseal the Anode Stack,
Bottom. Keep the stack
in the plastic tray.
3. Place the Anode Stack (in
the tray) on the blotting
surface. Align it with the
Gel Barriers on the right.
4. Place the pre-run gel(s)
on the transfer mem-
brane of the Anode
Stack.
Select button
Program Run Time
Program Voltage Default Run Time Run Time Limit Program Voltage Default Run Time Run Time Limit
P1 25 V 6 minutes 10 minutes P7 5 V 3 minutes 25 minutes
P2 23 V 6 minutes 11 minutes
P8
20 V for 2 minutes
23 V for 2 minutes
25 V for remainder
7 minutes 13 minutes
P3 20 V 7 minutes 13 minutes
P4 15 V 7 minutes 16 minutes
P5 10 V 7 minutes 25 minutes P9 20 V for 2 minutes
5 V for 3 minutes (x2) 8 minutes 8 minutes
P6 7.5 V 3 minutes 25 minutes
Selecting a Program
1. Press the power switch to turn ON the iBlot® Gel Transfer Device. The fan in the device
begins to run and digital display shows default parameters (P 3.0 7:00) or the last
program used.
2. Press the Select button to select the appropriate program. Use the Up/Down (+/-)
Buttons for changing the values to the recommended parameters listed above.
The iBlot® Gel Transfer Device is pre-programmed with 9 voltage programs listed below:
The recommended settings for running a Mini Transfer Stack with a Novex® mini-gel (1.0 or 1.5 mm thick), or a Regular Transfer
Stack with an E-PAGE™ 48 Gel, E-PAGE™ 96 Gel, Novex® midi-gel (1 mm thick), or 2 Novex® mini-gels (1.0 or 1.5 mm thick) is
program P3 for 7–8 minutes.
Using the iBlot® Gel Transfer Device with Blotting Roller
Instructions are provided below to assemble the iBlot® Gel Transfer Stack to perform blotting of mini-, midi-, or other gels using
the iBlot® Gel Transfer Device. Refer to the manual if you are using the iBlot® Gel Transfer Device with the de-bubbling roller.

Using the iBlot® Gel Transfer Device with Blotting Roller
©2011 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective
owners.
For technical support, email [e-mail].
For country-specific contact information, visit www.invitrogen.com.
1. Close the lid and secure
the latch. The red light
is on indicating a closed
circuit.
2. Ensure the correct
program and time are
selected.
3. Press the Start/Stop but-
ton. The red light chang-
es to green.
4. Current automatically
shuts off at the end of
each run. The end of
transfer is indicated by
beeping sounds, and
flashing red light and
digital display. Press and
release the Start/Stop
Button. The light turns to
a steady red.
5. Place the pre-soaked (in
deionized water) iBlot®
Filter Paper on the pre-
run gel and remove air
bubbles using the Blot-
ting Roller.
6. Unseal the Cathode
Stack, Top (shown here
for a Mini Stack). Discard
the red plastic tray.
7. Place the Cathode Stack,
Top over the pre-soaked
Filter paper with the
electrode side facing up
and aligned to the right
edge. Remove air bub-
bles using the Blotting
Roller.
8. Place the Disposable
Sponge with the metal
contact on the upper
right corner of the lid.
Proceed to Performing
Blotting, below.
Performing Blotting
Perform blotting within 15 minutes of assembling the stacks with the gel.
Disassembling the iBlot® Gel Transfer Device
1. Open the lid of the iBlot® Device.
2. Remove the iBlot® E-PAGE™ Tab (used for blotting E-PAGE™ gels only). Rinse the tab with deionized water and store in a dry
place for future use.
3. Discard the iBlot® Disposable Sponge and iBlot® Cathode Stack, Top.
4. Carefully remove and discard the gel and lter paper. Remove the transfer membrane from the stack and proceed with the
blocking procedure or stain the membrane.
5. Discard the iBlot® Anode stack, Bottom.
6. At this point, the iBlot® Gel Transfer Device is ready for another run (no cooling period is required). If you are not using the
device, turn off the power switch.
Downloading Upgrades
To download iBlot® Device rmware upgrades, go to www.invitrogen.com/iblot. Follow instructions on the page to download
the upgrades.
Limited Use Label License
This product is covered by Limited Use Label License 358: For Research Use Only: The purchase of this product conveys to the purchaser the
limited, non-transferable right to use the purchased amount of the product only to perform internal research for the sole benet of the purchaser.
No right to resell this product or any of its components is conveyed expressly, by implication, or by estoppel. This product is for internal
research purposes only and is not for use in commercial applications of any kind, including, without limitation, quality control and commercial
services such as reporting the results of purchaser’s activities for a fee or other form of consideration. For information on obtaining additional
rights, please contact [email protected] or Out Licensing, Life Technologies, 5791 Van Allen Way, Carlsbad, California 92008.