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Invitrogen™ iBind™ Solution Kit

Catalog No. SLF1020X4
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Description

Includes

Kit contains sufficient solution for 40 mini-membrane reactions.

Requires

iBind Western Device

For use with the iBind Western Device

The automated iBind Western Device performs every step in the Western processing workflow from blocking to washes to antibody incubations. The iBind solutions act as the blocking, washing, and dilution solutions in the iBind immunodetection process and have been optimized for ideal viscosity. Each solution is released from the iBind device wells to the iBind Card through sequential lateral flow (SLF); the solutions are then wicked towards the stack region of the card. The glass fiber matrix of the card allows for homogenous and consistent flow of the solutions to the membrane, increasing the antigen-antibody interaction.

  • Optimized solutions with ideal viscosity for blocking, washing, and antibody dilution
  • Long shelf life—store iBind Solutions Kit for 12 months at 4°C
  • Better antigen-antibody interaction—solutions flow between the iBind Card and membrane for maximum interaction between protein and antibody
  • Brings automation and simplicity to the Western protocol

Proteins, Expression, Isolation and Analysis, Western Blotting

Specifications

Content And Storage • iBind 5X buffer (240 mL)
• iBind 100X additive (8 x 1.5 mL)

Store at 2–8°C.
Detection Method Colorimetric, Chromogenic, Chemiluminescence
For Use With (Application) Western Blot
For Use With (Equipment) iBind Western Device
Membrane Compatibility Nitrocellulose, PVDF
Product Line iBind
Throughput 1 Midi Blot, 6 Vertically-Cut Strip Blots, 2 Mini Blots/Card
Type Solution Kit
Includes iBind 5X buffer (240 mL); iBind 100X additive (8 x 1.5 mL)
No. of Reactions 10 mini membrane reactions
Quantity 4 kits
Shipping Condition Qualified for Ambient or Wet Ice
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Frequently Asked Questions (FAQs)

What procedure should I follow when stripping and reprobing my blot processed using the iBind Western Device or iBind Flex Western Device?

To strip and reprobe your blot processed using the iBind Western Device or iBind Flex Western Device, first strip the blot outside the system using your preferred protocol. After the stripping protocol, wash the blot in distilled water for 5 min. Reblocking is unnecessary as the iBind Solution will perform this step during reprobing. Finally, reprobe with new antibodies using the iBind Western Device or iBind Flex Western Device, starting with the membrane preparation step.

Can I multiplex antibodies using the iBind Western Device or iBind Flex Western Device?

Yes, for each midi blot, mini blot, or vertically cut strip, prepare appropriate primary antibodies together in a single tube of iBind FD/ iBind Flex FD Solution, Prepare appropriate secondary antibodies together in a different tube of iBind FD/ iBind Flex FD Solution. Do not mix primary and secondary antibodies prior to loading in the device. All primary and secondary antibodies should be diluted according to the recommendations outlined in the user manual or quick reference card. When detecting several targets of the same species, the same concentration of secondary antibody can be used as when detecting one target. There is no need to increase the concentration for each additional target. Load multiplexed antibodies into the device as normal.
Caution: Consider cross-reactivity of secondary antibodies when multiplexing (e.g., rabbit anti-goat IgG and goat anti-mouse IgG are likely to cross-react).

Can I use the iBind Western Device or iBind Flex Western Device with Alexa Fluor Plus conjugated secondary antibodies?

Yes, both the iBind Western Device and iBind Flex Western Device work well with Alexa Fluor Plus conjugated secondary antibodies.

With the iBind Western Device and iBind Flex Western Device, how do I know if I am rolling the membrane firmly enough?

When firmly rolling, the iBind/iBind Flex Card will slightly dip in the membrane area, causing the top and bottom of the card to lift. See the iBind/iBind Flex user manual for additional details.
Is there any difference between the iBind Flex Card and the original iBind Card?

Besides being ~1.7 times as wide, the iBind Flex Card also includes alignment lines printed on the back that appear when solution is added to the card to help in proper horizontal placement of multiple mini or vertically cut strip blots.
Is there any difference between the iBind Flex Solution Kits and the original iBind Solution Kits?

The formulations are identical for both the standard and Fluorescent Detection (FD) solutions. The only difference is in the amount supplied, so the iBind Flex Solution Kits are sufficient for 10 midi, 20 mini, or 60 vertically cut strip blots.
For the iBind Western System, do you have a protocol for detecting the signal using the LI-COR Odyssey Imaging System?

Please use the protocol mentioned on Page 13 of the manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/ibind_man.pdf) when using the iBind Western System in conjunction with the LI-COR Odyssey Imaging System. We do not recommend using the LI-COR solution.
When is it necessary to pre-activate a PVDF membrane with methanol for use in the iBind Western Device?

If the PVDF membrane has dried up, it is important to pre-activate it with 100% methanol and then rinse in distilled water before immersing it in iBind Solution/iBind Fluorescent Detection (FD) Solution. It is not necessary to activate a PVDF membrane that has just come out of the transfer and moved into iBind Solution/iBind Fluorescent Detection (FD) Solution.
What are your recommendations for cleaning and maintenance of the iBind Western device?

Rinse the iBind Western Device under running water after each use and allow the device to dry before additional usage. No use of detergent is recommended as any residual detergent may interfere with results. To maximize the life of the springs in the iBind Western Device, store the device with latch unlocked, and the lid open as shown on Page 23 of the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf).
What is the recommended amount of time to immerse the membrane in iBind Solution/ iBind Fluorescent Detection (FD) Solution prior to assembly in the iBind Western device?

It is sufficient to block the membrane in iBind Solution/ iBind Fluorescent Detection (FD) Solution for the time prior to making antibody dilutions. It is not necessary to block the membrane overnight in the blocking solution. The membrane will be subjected to additional blocking in the iBind Solution/ iBind Fluorescent Detection (FD) Solution before primary antibody reaches the membrane.
The iBind Western System protocol recommends trimming the membrane to 9 cm x 9 cm. Can I use a membrane smaller than that?

Yes, membranes that are smaller than 9 cm x 9 cm can be used with the iBind Western device. It is recommended to place the strips or smaller blots centrally. When using larger membranes, care must be taken to ensure that no part of the membrane is in contact with the stack on the iBind card.
Can I use my own blocking reagent or antibody incubation solution with the iBind Western System?

We do not recommend using your own solutions. The iBind Solution and iBind Fluorescent Detection (FD) Solution have specific viscosity and are optimized for the sequential lateral flow that is the principle of the iBind Western device. We cannot guarantee the performance with any other solutions.
What are the components of the original iBind Western System and which components do I need to purchase separately?

The original iBind Western System (Cat. No. SLF1000) consists of:
- iBind Western Device
- iBind Blotting Roller
- iBind Window Cover

You would need to purchase the following items separately:
- iBind Cards (Cat. No. SLF1010)
- iBind Solution Kit (Cat. No. SLF1020) for preparing blocking, dilution, and washing buffers for chemiluminescent or chromogenic detection
- iBind Fluorescent Detection Solution Kit (Cat. No. SLF1019) for preparing blocking, dilution, and washing buffers for fluorescent detection
- Invitrogen AP Mouse Chemiluminescent Detection Kit (Cat. No. SLF1021) or Invitrogen AP Rabbit Chemiluminescent Detection Kit (Cat. No. SLF1022)
How do iBind Western Systems work?

iBind Western Systems are benchtop devices that automate immunodetection steps. These devices use no external power source, and rely on mechanical pressure to generate sequential lateral flow (SLF) of immunodetection reagents to perform hands-free blocking, antibody binding, and washes for western detection workflows.

Two iBind Western Systems are available:
i) containing the original iBind Western Device, which accommodates one mini blot at a time
ii) containing the iBind Flex Western Device, which accommodates one or two mini blots, one midi blot, or up to six vertically cut membrane strips (for separate antibodies) at a time.

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