Product FAQ

Are the iBlot Transfer Stacks compatible with Li-COR detection?

Answer

Yes, the iBlot Transfer Stacks are compatible with Li-COR detection.

Answer Id: E11395

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Product FAQ

Do you still offer the iBlot Gel Transfer Device?

Answer

The iBlot Gel Transfer Device has been discontinued and we have launched the iBlot 2 Gel Transfer Device, that is an improved version. The iBlot Transfer Stacks, Western Detection Stacks and DNA Transfer Stacks are still available for purchase and are to be used exclusively with the original iBlot Gel Transfer Device, and are not compatible with the new iBlot 2 Gel Transfer Device (http://www.thermofisher.com/order/catalog/product/IB21001). Only iBlot 2 consumables are to be used with the iBlot 2 Gel Transfer Device.

Answer Id: E11380

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Manual / Product Insert

iBlot Western Detection Kits

Version: Rev date: 18 June 2010
Catalog #
  • CB711001(Discontinued)
  • CB711002(Discontinued)
  • CB721001(Discontinued)
  • CB721002(Discontinued)
  • CB731001(Discontinued)
  • CB731002(Discontinued)
  • CB741001(Discontinued)
  • CB741002(Discontinued)
  • IB701001(Discontinued)
  • IB701002(Discontinued)
  • IB711001(Discontinued)
  • IB711002(Discontinued)
  • IB721001(Discontinued)
  • IB721002(Discontinued)
  • IB731001(Discontinued)
  • IB731002(Discontinued)
  • IB741001(Discontinued)
  • IB741002(Discontinued)

Manual / Product Insert

iBlot DNA Transfer Stack QRC

Version: Version A; 6 August 2008
Catalog #
  • IB801001
  • IB1001EU(Discontinued)
  • IB1001UK(Discontinued)
  • IB1001(Discontinued)

Product Literature

Application Note: Transfer of high molecular weight proteins using the iBlot 2 Gel Transfer Device

Product FAQ

I used the iBind Flex Western system and am getting high background. What should I do?

Answer

Here are possible causes and solutions:

- Membrane not completely wet: Follow instructions for prewetting the membrane. Use an incubation dish which is small enough to allow thorough coverage of membrane to prevent drying out. Note: If PVDF membrane is being used, we recommend making sure that it is activated with methanol, especially if it has dried up. It is not necessary to activate a PVDF membrane that has just come out of the transfer and moved into 1X iBind Flex Solution/ iBind Flex FD Solution.
- Membrane is contaminated: Use only clean, new membranes. Wear clean gloves at all times and use forceps when handling membranes.
- Film overexposed or became wet during exposure: Decrease exposure time or allow signal to further decay. Prevent leakage by encasing membrane in transparency film and blotting excess substrate from edges before exposure.
- Solutions or incubation tray are contaminated: Use clean glassware and purified water to prepare solutions. Replace or clean the tray thoroughly with a glassware-cleaning detergent. Rinse thoroughly with purified water. Wear clean gloves at all times.
- Concentrated primary antibody used: Follow the supplier's recommended dilution or determine the optimum concentration by dot blotting.
- Incorrect chemiluminescent substrate used for PVDF: Make sure CDP-Star reagent without enhancer is used.
- Blot is overdeveloped: Follow recommended developing time or remove blot from substrate when signal-to-noise ratio is acceptable.
- Ink used to label membrane: Any labeling of the membrane with ink should be limited to the low MW region of the blot.
- Improper preparation of iBind Flex Solution/ iBind Flex FD Solution: Prepare 1X iBind Flex Solution/ iBind Flex FD Solution as directed in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Improper application of solutions to iBind Flex Wells: Add the appropriate solutions for each well in the correct numerical order as specified in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_flex_man.pdf).
- Blot improperly placed on iBind Flex Card: 1) Place the membrane in the designated Membrane Region on the iBind Flex Card. 2) The protein side of the blot should be in contact with the iBind Flex Card. 3) The low MW regions should be closest to the Stack. 4) The membrane should not be in contact with the Stack.
- Card stack wet prior to run: Ensure that 10 mL of 1X iBind Flex/iBind Flex FD Solution is added to the flow region of the card. Avoid adding the solution to the stack.

Here are some additional tips to reduce background:

- If iBlot PVDF stacks are used, check that they have not expired as background increases with age. Using the iBlot 2 PVDF stacks will help in reducing the background.
- Make sure that the blot is rinsed in distilled water prior to adding the substrate. Do not rinse in PBS or TBS.

Answer Id: E11312

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Product FAQ

I used the iBind Western system and am getting high background. What should I do?

Answer

Here are possible causes and solutions:

- Membrane not completely wet: Follow instructions for prewetting the membrane. Use an incubation dish which is small enough to allow thorough coverage of membrane to prevent drying out.
Note: If PVDF membrane is being used, we recommend making sure that it is activated with methanol, especially if it has dried up. It is not necessary to activate a PVDF membrane that has just come out of the transfer and moved into 1X iBind Solution/ iBind FD Solution.
- Membrane is contaminated: Use only clean, new membranes. Wear clean gloves at all times and use forceps when handling membranes.
- Film overexposed or became wet during exposure: Decrease exposure time or allow signal to further decay. Prevent leakage by encasing membrane in transparency film and blotting excess substrate from edges before exposure.
- Solutions or incubation tray are contaminated: Use clean glassware and purified water to prepare solutions. Replace or clean the tray thoroughly with a glassware-cleaning detergent. Rinse thoroughly with purified water. Wear clean gloves at all times.
- Concentrated primary antibody used: Follow the instructions provided in the manual ((https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf)) to dilute the primary antibody or determine the optimum concentration by dot blotting.
- Incorrect chemiluminescent substrate used for PVDF: Make sure CDP-Star reagent without enhancer is used.
- Blot is overdeveloped: Follow recommended developing time or remove blot from substrate when signal-to-noise ratio is acceptable.
- Ink used to label membrane: Any labeling of the membrane with ink should be limited to the low MW region of the blot.
- Improper preparation of iBind Solution/ iBind FD Solution: Prepare 1X iBind Solution/ iBind FD Solution as directed in the manual.
- Improper application of solutions to iBind Wells: Add the appropriate solutions for each well in the correct numerical order as specified on Page 18 in the manual (https://tools.thermofisher.com/content/sfs/manuals/ibind_man.pdf).
- Blot improperly placed on iBind Card: 1) Place the membrane in the designated Membrane Region on the iBind Card. 2) The protein side of the blot should be in contact with the iBind Card. 3) The low MW regions should be closest to the Stack. 4) The membrane should not be in contact with the Stack.

Here are some additional tips to reduce background:

- If iBlot PVDF stacks are used, check that they have not expired as background increases with age. Using the iBlot 2 PVDF stacks will help in reducing the background.
- Make sure that the blot is rinsed in distilled water prior to adding the substrate. Do not rinse in PBS or TBS.

Answer Id: E11306

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Product Literature

BioProbes 78 Journal of Cell Biology Applications

Product Literature

Technical handbook: Protein gel electrophoresis

Manual / Product Insert

Novex Chromogenic Substrates

Version: 28 May 2008

Product Literature

Lab consumables catalog 2017[JA]

Product Literature

Life in the Lab - Stay Curious (North America)

Product Literature

フライヤー: iBlot 2 ドライブロッティングシステム

Product Literature

ラボプロダクツ総合カタログ2017