Life technologies Invitrogen iBlot Quick Reference Card

iBlot dry Blotting System
®
catalog numbers: iB1001, iB1001eu, iB1001uk
Rev. date: 23 february 2011
Instructions for using the iBlot
Transfer Stacks is described below. For detailed instructions, including instructions for transfer of proteins from E-PAGE
transfer of DNA from agarose gels or polyacrylamide gels to iBlot
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
blotting E-PAGE , 1 midi-, or 2 mini-gels. Use iBlot
™
• 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.

Selecting a Program

1. Press the power switch to turn ON the iBlot
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:
®
Program Voltage Default Run Time
P1 25 V
P2 23 V
P3 20 V
P4 15 V
P5 10 V
P6 7.5 V
The recommended settings for running a Mini Transfer Stack with a Novex
Stack with an E-PAGE 48 Gel, E-PAGE
™
program P3 for 7–8 minutes.
using the iBlot Gel Transfer device with Blotting Roller
®
Instructions are provided below to assemble the iBlot
the iBlot Gel Transfer Device. Refer to the manual if you are using the iBlot
®
1. Open the lid of the device.
intended use: For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Gel Transfer Device to perform dry blotting of proteins from mini- or midi-gels with iBlot
®
® Gel Transfer Stacks to fit your gel size. Use the iBlot
Gel Transfer Device. The fan in the device
®
Run Time Limit
6 minutes 10 minutes
6 minutes 11 minutes
7 minutes 13 minutes
7 minutes 16 minutes
7 minutes 25 minutes
3 minutes 25 minutes
96 Gel, Novex
™
®
2. Unseal the Anode Stack,
Bottom. Keep the stack
in the plastic tray.
Part number 25-0912
DNA Transfer Stacks, and performing western detection with
®
Gel Transfer Stacks, Mini for blotting 1 mini-gel.
®
Program Voltage
P7 5 V
20 V for 2 minutes
23 V for 2 minutes
P8
25 V for remainder
20 V for 2 minutes
P9
5 V for 3 minutes (x2)
mini-gel (1.0 or 1.5 mm thick), or a Regular Transfer
®
midi-gel (1 mm thick), or 2 Novex
®
Gel Transfer Stack to perform blotting of mini-, midi-, or other gels using
Gel Transfer Device with the de-bubbling roller.
®
3. Place the Anode Stack (in
the tray) on the blotting
surface. Align it with the
Gel Barriers on the right.
For technical support, email tech_support@invitrogen.com.
For country-specific contact information, visit www.invitrogen.com.
MAN0003281
® Gel Transfer Stacks, Regular for
Program Run Time Select button
Default Run Time Run Time Limit
3 minutes 25 minutes
7 minutes 13 minutes
8 minutes 8 minutes
mini-gels (1.0 or 1.5 mm thick) is
®
4. Place the pre-run gel(s)
on the transfer mem-
brane of the Anode
Stack.
Quick
Quick
RefeRence
RefeRence
Guide
caRd
Gel
®
™ gels,