Indigo Biosciences INDIGO BIOSCIENCES INC Reporter Assay Kit, Human ADRA2A, 1 x 96-well format

Catalog No. NC3978873 Supplier Indigo Biosciences Supplier No. IB34001

Frequently Asked Questions (FAQs)

I used PVDF Gel Transfer Stacks with the iBlot 3 Western Blot Transfer System, and my proteins did not transfer to the membrane. What happened?

This is likely due to the PVDF membrane being dry or partially dry. Regions where PVDF membranes are dry appear whiter than places where the membrane is wet. Remove the membrane and reactivate in 100% methanol, and rinse in water before reapplying to the transfer stack.

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.

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.

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.

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.

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.

Are the copper electrodes in the iBlot 3 Transfer Stacks recyclable?

To maximize the recovery of the copper from the mesh electrodes used in the iBlot 3 Transfer Stacks, we have established an agreement with a recycling center in the United States. In order to prepare the copper electrodes for recycling, please follow the instructions listed here.

I want to perform western transfer with mini gels (8 x 8 cm), but I do not have iBlot 3 Transfer Stacks (Mini). Can I use iBlot 3 Transfer Stacks (Regular) to transfer my mini gels?

It is best not to transfer a single mini gel on a regular-sized transfer stack. Although in most cases, the transfer will be fine, empty spaces on the transfer stack that are not in direct contact with the gel could potentially cause distortions across the whole surface of the membrane, including the portion in contact with the gel. It is best to have more than 50% of a membrane in contact with the gel, if possible. Our recommendation is that you transfer 2 mini gels using a Regular transfer stack.

In the iBlot 3 Top Stack, I see a white sheet beneath the transfer gel layer. What purpose does it serve?

This is the plastic separator that separates the Top Stack from the Bottom Stack. Right before assembling the transfer stack, this separator needs to be removed from the Top Stack and discarded.

Is it okay to freeze the iBlot 3 Transfer Stacks?

The iBlot 3 Transfer Stacks should not be frozen as they contain a gel matrix that will be damaged by freezing.

Can I use the iBlot 3 Transfer Stacks after the expiration date?

No, we do not recommend using the iBlot 3 Transfer Stacks after the expiration date. Use of transfer stacks after the expiration date can lead to inferior results.

What is the recommended storage condition for the iBlot 3 Transfer Stacks?

We recommend storing the iBlot 3 Transfer Stacks at room temperature.

What are the different kinds of iBlot 3 Transfer Stacks that you offer and are they available separately?

We offer iBlot 3 Transfer Stacks with integrated nitrocellulose or PVDF membranes for transferring proteins from gels onto nitrocellulose or PVDF membranes, respectively (western blotting). These transfer stacks are available in Midi and Mini format and can also be purchased separate from the iBlot 3 Western Blot Transfer Device.

Can I trim the Bottom Stack of my iBlot 3 Transfer Stack to fit my separating gel?

No, the iBlot 3 Transfer Stack should not be trimmed to fit your gel size.

Is the PVDF membrane in the iBlot 3 PVDF Transfer Stack pre-activated?

Yes, the PVDF membrane is pre-activated and ready for use.

Can I use the iBlot 3 Transfer Stacks and/or iBlot 3 Absorbent Pad more than once?

No, the iBlot 3 Transfer Stacks contain a finite number of ions to drive the transfer and are depleted after a single use. We also recommend discarding the iBlot 3 Absorbent Pad after every use.

Is there a limitation on the thickness of gels that can be used with the iBlot 3 Western Blot Transfer Device?

The iBlot 3 Western Blot Transfer Device is compatible with gels with a thickness of ≤1.5 mm.

Are the iBlot 3 Transfer Stacks compatible with the original iBlot and iBlot 2 transfer devices?

Unfortunately, the iBlot 3 Transfer Stacks are not compatible with the original iBlot and iBlot 2 transfer devices. The iBlot 3 Transfer Stacks are only compatible with the iBlot 3 Western Blot Transfer Device.