After transferring using the iBlot 3 Western Blot Transfer System, I noticed that the Bottom Stack transfer gel melted to a viscous blue solution. Why is this?
This happens if the membrane is trimmed to fit the gel size resulting in direct contact between the Top and Bottom transfer stacks. This can be avoided by maintaining the membrane size identical to the transfer stack. Transfer quality is not affected by smaller gel size compared to the membrane.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
What is the Blotting Roller that is provided with the iBlot 3 Western Blot Transfer System?
The Blotting Roller is a plastic roller attached to a stainless steel handle. It is used to remove any air bubbles between the gel and blotting membrane during the assembly of the transfer stacks and gel.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Can I cut the membrane in the iBlot 3 Transfer Stacks to fit my separating gel?
No, we recommend maintaining the membrane size identical to the transfer stacks. This helps ensure that there is no direct contact between the Top and Bottom transfer stacks.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Are the PVDF and nitrocellulose membranes in the iBlot 3 Transfer Stacks compatible with fluorescent antibodies or membrane staining (e.g., LI-COR, Ponceau S)?
The PVDF and nitrocellulose membranes in the iBlot 3 Transfer Stacks are compatible with all commonly used detection methods such as staining, immunodetection, fluorescence, etc.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.
Can I replace the membrane in the transfer stack with my preferred membrane when using the iBlot 3 Western Blot System?
Yes, it is possible to replace the membrane provided in your transfer stack with any membrane that is compatible with western blotting. To do this, cut the alternative membrane to match the size of your gel, and wet the membrane. Then either place the alternative membrane on top of the integrated membrane, or carefully remove the integrated membrane from the gel matrix with forceps and replace it with the new membrane. Note that Thermo Fisher Scientific only supports the use of transfer stacks when they are used with the provided instructions.
Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.