Invitrogen iBlot 2 Gel Transfer Device

Find answers to common questions about the iBlot 2 Gel Transfer Device, download the latest firmware, and register your device.

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Q: Is the iBlot 2 Gel transfer device compatible with original iBlot Transfer Stacks?
A: No. The original iBlot Transfer stacks are not compatible.

Q: Sometimes there is green discoloration on the blot around the gel. What is this, and does it affect the results?
A: The green discoloration is copper deposits from the transfer stack, and it does not affect the results. To minimize this effect, shake excess water off the filter paper and buffer from the gel before placing each on the stack. The current formulation of stacks minimizes the green discoloration.

Q: What is the shelf life of the iBlot 2 Transfer Stack?
A: The transfer stacks are good for 9 months. Expiration date is printed on each consumable.

Q: Can I use the stacks after the expiration date?
A: It is not recommended to use the stacks after the expiration date has passed. The results may be inferior.

Q: Can the iBlot 2 be used for transfer of low- and high- molecular weight proteins?
A: Yes. Optimization to the transfer protocol may be needed for some proteins. Suggestions are listed in the manual for transfering low and high molecular weight proteins.

Q: Can I create my own transfer conditions?
A: Yes, the device allows programming and saving custom methods.

Q: I want to conduct western transfer with mini gels (8 x 8 cm), but I don’t have iBlot Transfer Stacks (Mini). Can I use iBlot Transfer Stacks (Regular) to transfer my mini gels?
A: 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 a 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.

Q: Can I preform multiple transfers, with a new transfer starting immediately after the previous one?
A: Yes, the device is designed to do so with no impact on performance.

Q: Can I use the stack and /or absorbent pad more than once?
A: No. The transfer stacks have a finite amount of ions to drive the transfer and are depleted after a single use.

Q: Are the iBlot 2 PVDF and NC membranes compatible with fluorescence or membrane staining?
A: The membranes are compatible with all commonly used detection methods such as staining, chemiluminescence, chromogenic and fluorescence.

Q: Is it possible to substitute the membrane with my specialized membrane?
A: 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.

Q: Is the PVDF membrane pre-activated?
A: Yes, the membrane is pre-activated and ready for use without having to perform any pre-treatment with alcohol.

Q: Can I cut the membrane before transfer to fit my gel?
A: No, it is important to maintain the membrane size identical to the transfer stack. This helps ensure that there is no direct contact between the top and bottom transfer stacks.

Q: Can I cut the stack to fit my gel?
A: No, do not trim the iBlot 2 stacks.

Q: How can I get better transfer of high molecular weight proteins?
A: Proteins larger than ~150 kDa migrate more slowly than smaller proteins. Therefore, more time is required to transfer them from gel to membrane. We recommend extending the transfer time by 8–10 minutes for optimal transfer of proteins >150 kDa. To enhance transfer efficiency, we also recommend adding an equilibration (gel-soaking) step between electrophoresis and western transfer and using NuPAGE Novex 3-8% Tris-acetate gels for electrophoresis. Please refer to the application note in the application note tab for more details.

Q: What causes empty spots on my membrane after transfer?
A: The iBlot 2 Dry Blotting System is similar to conventional transfer methods in that air bubbles between the gel and the membrane will prevent protein transfer. Be sure to remove all air bubbles between the gel and membrane before starting the transfer, using the blotting roller supplied.

Q: Can I still buy consumables for the original iBlot Gel Transfer Device?
A: Yes, they are still available. You can find them listed here.

Q: Is the iBlot 2 Gel Transfer Device compatible with original iBlot Transfer Stacks?
A: No. The iBlot Transfer Stacks do not work in the iBlot 2 Gel Transfer Device.

Q: Is there a limitation on the thickness of gels that can be used with the iBlot Dry Blotting System?
A: The iBlot Dry Blotting System has been tested to efficiently transfer protein from gels ranging in thickness from 1 mm to 3 mm. Gels thicker than 3 mm have not been tested.

Q: How can we be more environmentally responsible?
A: The plastic in the iBlot Dry Blotting System stack packaging is polyethylene terephthalate (PET) and can be recycled.

Q: Are the copper electrodes in the transfer stacks recyclable?
A: No. The electrodes are copper-coated nylon, and the amount of copper left in the electrode after a transfer is rather small. There is 0.6 grams of copper per sheet before transfer, and even less after transfer.

Q: What is the shelf life of both the nitrocellulose and PVDF iBlot Transfer Stacks?
A: The minimum guaranteed shelf life for iBlot Transfer Stacks is 2 months. Depending on when you purchase the transfer stack, shelf life will be 2–8 months.

Q: Does the iBlot Dry Blotting System work with native or native-blue gels?
A: Yes, we have an application note available, titled, "Western Blotting NativePAGE Novex Bis-Tris gels Using the iBlot 7-Minute Blotting System". To download the pdf, see Application notes tab on this page.

Q: Is the iBlot 2 Gel transfer device compatible with original iBlot Transfer Stacks?
A: No. The original iBlot Transfer stacks are not compatible.

Q: Sometimes there is green discoloration on the blot around the gel. What is this, and does it affect the results?
A: The green discoloration is copper deposits from the transfer stack, and it does not affect the results. To minimize this effect, shake excess water off the filter paper and buffer from the gel before placing each on the stack. The current formulation of stacks minimizes the green discoloration.

Q: What is the shelf life of the iBlot 2 Transfer Stack?
A: The transfer stacks are good for 9 months. Expiration date is printed on each consumable.

Q: Can I use the stacks after the expiration date?
A: It is not recommended to use the stacks after the expiration date has passed. The results may be inferior.

Q: Can the iBlot 2 be used for transfer of low- and high- molecular weight proteins?
A: Yes. Optimization to the transfer protocol may be needed for some proteins. Suggestions are listed in the manual for transfering low and high molecular weight proteins.

Q: Can I create my own transfer conditions?
A: Yes, the device allows programming and saving custom methods.

Q: I want to conduct western transfer with mini gels (8 x 8 cm), but I don’t have iBlot Transfer Stacks (Mini). Can I use iBlot Transfer Stacks (Regular) to transfer my mini gels?
A: 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 a 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.

Q: Can I preform multiple transfers, with a new transfer starting immediately after the previous one?
A: Yes, the device is designed to do so with no impact on performance.

Q: Can I use the stack and /or absorbent pad more than once?
A: No. The transfer stacks have a finite amount of ions to drive the transfer and are depleted after a single use.

Q: Are the iBlot 2 PVDF and NC membranes compatible with fluorescence or membrane staining?
A: The membranes are compatible with all commonly used detection methods such as staining, chemiluminescence, chromogenic and fluorescence.

Q: Is it possible to substitute the membrane with my specialized membrane?
A: 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.

Q: Is the PVDF membrane pre-activated?
A: Yes, the membrane is pre-activated and ready for use without having to perform any pre-treatment with alcohol.

Q: Can I cut the membrane before transfer to fit my gel?
A: No, it is important to maintain the membrane size identical to the transfer stack. This helps ensure that there is no direct contact between the top and bottom transfer stacks.

Q: Can I cut the stack to fit my gel?
A: No, do not trim the iBlot 2 stacks.

Q: How can I get better transfer of high molecular weight proteins?
A: Proteins larger than ~150 kDa migrate more slowly than smaller proteins. Therefore, more time is required to transfer them from gel to membrane. We recommend extending the transfer time by 8–10 minutes for optimal transfer of proteins >150 kDa. To enhance transfer efficiency, we also recommend adding an equilibration (gel-soaking) step between electrophoresis and western transfer and using NuPAGE Novex 3-8% Tris-acetate gels for electrophoresis. Please refer to the application note in the application note tab for more details.

Q: What causes empty spots on my membrane after transfer?
A: The iBlot 2 Dry Blotting System is similar to conventional transfer methods in that air bubbles between the gel and the membrane will prevent protein transfer. Be sure to remove all air bubbles between the gel and membrane before starting the transfer, using the blotting roller supplied.

Q: Can I still buy consumables for the original iBlot Gel Transfer Device?
A: Yes, they are still available. You can find them listed here.

Q: Is the iBlot 2 Gel Transfer Device compatible with original iBlot Transfer Stacks?
A: No. The iBlot Transfer Stacks do not work in the iBlot 2 Gel Transfer Device.

Q: Is there a limitation on the thickness of gels that can be used with the iBlot Dry Blotting System?
A: The iBlot Dry Blotting System has been tested to efficiently transfer protein from gels ranging in thickness from 1 mm to 3 mm. Gels thicker than 3 mm have not been tested.

Q: How can we be more environmentally responsible?
A: The plastic in the iBlot Dry Blotting System stack packaging is polyethylene terephthalate (PET) and can be recycled.

Q: Are the copper electrodes in the transfer stacks recyclable?
A: No. The electrodes are copper-coated nylon, and the amount of copper left in the electrode after a transfer is rather small. There is 0.6 grams of copper per sheet before transfer, and even less after transfer.

Q: What is the shelf life of both the nitrocellulose and PVDF iBlot Transfer Stacks?
A: The minimum guaranteed shelf life for iBlot Transfer Stacks is 2 months. Depending on when you purchase the transfer stack, shelf life will be 2–8 months.

Q: Does the iBlot Dry Blotting System work with native or native-blue gels?
A: Yes, we have an application note available, titled, "Western Blotting NativePAGE Novex Bis-Tris gels Using the iBlot 7-Minute Blotting System". To download the pdf, see Application notes tab on this page.

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