Quick
RefeRence
Guide
iBlot
®
dry Blotting System
catalog numbers: iB1001, iB1001eu, iB1001uk
Rev. date: 23 february 2011
Quick
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 [email protected].
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 fit 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.
