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Invitrogen™ iBlot™ DNA Transfer Stack

Catalog No. IB801001
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IB801001 DNA Transfer Stack
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Catalog No. IB801001 Supplier Invitrogen™ Supplier No. IB801001

Please call Customer Service at 1-800-234-7437 or send an email to help@thermofisher.com for assistance.

Description

Includes

Each iBlot DNA Transfer Stack contains sponges, cathode and anode electrodes and a nylon membrane, preassembled and primed with a proprietary formulation for optimum transfer of DNA from the gel to the membrane. Transfer stacks are offered in 10-pack sizes.

Quick and efficient system for transferring DNA from agarose or polyacrylamide gels in seven minutes

Unlike capillary, vacuum, or electroblotting, dry blotting requires no additional buffers or liquids resulting in less result variability.

  • Time savings—transfer DNA in 7 minutes instead of 16 hours
  • Reproducibility—consistent blot preparation and running conditions
  • Convenience—avoid messy buffers and laborious prep work
  • Consists of a Gel Transfer Device and Transfer Stacks
  • Transfer stacks are preassembled and intended for single use only
  • Pore size: 0.2μm
  • Gel size: Midi (8 x 13cm)

DNA and RNA Purification and Analysis, Nucleic Acid Gel Electrophoresis and Blotting, Southern Blotting

Order Info

Shipping Condition: Room temperature

Specifications

Content And Storage Each iBlot™ DNA Transfer Stack contains sponges, cathode and anode electrodes and a nylon membrane, pre-assembled and primed with a proprietary formulation for optimum transfer of DNA from the gel to the membrane. Transfer stacks are offered in 10-pak sizes. Store iBlot™ DNA Transfer Stacks at room temperature. This product is guaranteed stable for 2 months when properly stored.
For Use With (Equipment) iBlot
Gel Compatibility Polyacrylamide Gel, Agarose Gel
Gel Size Midi (8 x 13 cm)
Membrane Compatibility Nylon
Mode of Transfer Dry
Product Line iBlot
Type DNA Transfer Stack
Membrane Nylon Membrane
Pore Size 0.2 micron
Quantity 10 stacks
Shipping Condition Room Temperature
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Frequently Asked Questions (FAQs)

Following the release of the iBlot 2 Gel Transfer Device, are consumables for the original iBlot Gel Transfer Device still available?

Yes, all original iBlot stacks are still available for purchase. You can find them in the original iBlot Gel Transfer Device Reagents and Resources (http://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/western-blotting/transfer-proteins-western-blot/iblot-dry-blotting-system/reagents-resources-original-iblot-gel-transfer-device.html). However, please note that they are not compatible with the iBlot 2 Gel transfer Device.
Is the iBlot 2 Gel Transfer Device compatible with original iBlot Transfer Stacks?

No. Do not use iBlot Transfer Stacks in the iBlot 2 Gel Transfer Device, and do not mix components between iBlot Transfer Stacks and iBlot 2 Transfer Stacks.
How can I get better transfer of high-molecular weight proteins?

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 to 10 minutes for optimal transfer of proteins >150 kDa using the iBlot Dry Blotting System.

To enhance transfer efficiency, we also recommend adding an equilibration (gel-soaking) step between electrophoresis and western transfer and using NuPAGE Invitrogen 3 - 8% Tris-acetate gels for electrophoresis. We have an application note available, titled “Transferring Large and Small Proteins Using the iBlot Dry Blotting System”. To download the PDF, see the iBlot Reagents Resources page. (http://www.thermofisher.com/us/en/home/life-science/protein-expression-and-analysis/western-blotting/western-blot-transfer/iblot-dry-blotting-system/original-iblot-resources.html)

Occasionally my western blots have high background. What do you recommend?

This may be a result of insufficient blocking or nonspecific binding. We suggest trying our WesternBreeze Blocker/Diluent (Cat. No. WB7050). We have been using it with good success. Additionally, you should optimize primary and secondary antibody concentrations as generally recommended for any new blotting technique. Many cases of high background can be resolved by further diluting one or both antibody preparations.
Is it possible to substitute the membrane from the iBlot Transfer Stack with my specialized membrane?

In theory, you can replace the membrane provided in your iBlot 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 we only support the use of iBlot Transfer Stacks when they are used with the provided instructions.
What is the shelf life of nitrocellulose and PVDF iBlot Transfer Stacks?

The minimum guaranteed shelf life of iBlot Transfer Stacks is 2 months. Depending on when you purchase the transfer stack, shelf life will be 2–8 months. The expiration date is printed on the package for each stack.
Can iBlot Transfer Stacks be used more than once?

No. The transfer stacks have a finite amount of ions to drive the transfer and are depleted after a single use.
Do the PVDF iBlot Transfer Stacks (0.2 µm, non-autofluorescing) require activation prior to use?

No. The PVDF membrane comes preactivated. You just need to open the transfer stack with membrane, place the separation gel on top of the membrane, and apply one layer of moistened filter paper to run (the same as with the nitrocellulose stacks).
Sometimes there is green discoloration on the blot around the gel when using the iBlot Dry Blotting System. What is this, and does it affect the results?

The green discoloration is copper deposits from the iBlot Transfer Stacks, 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.
How can we be more environmentally responsible in regards to the packaging for the iBlot Dry Blotting System?

The plastic in the iBlot Dry Blotting System stack packaging is polyethylene terephthalate (PET) and can be recycled.
I want to conduct western transfer with mini gels (8 x 8 cm), but I do not have iBlot Transfer Stacks (Mini). Can I use iBlot 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 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.

For Research Use Only. Not for use in diagnostic procedures.