Product FAQ

I transferred my proteins using the iBlot Dry Blotting System but the high-molecular weight proteins remained in the gel indicated by staining of the gel after transfer. Can you help me troubleshoot?

Answer

This could happen if an incorrect voltage Method was used or if inappropriate transfer conditions were used. Make sure that the voltage Method and run time used is correct, based on the gel type, as described on page 13 in the manual.

For mini or midi gels:
- Perform an ethanol equilibration step as described on page 28 in the manual to improve transfer.
- Use a lower gel percentage to separate the high-molecular weight proteins.
- Increase the transfer time in 30-second increments.

For E-PAGE gels:
- Increase the transfer time in 30-second increments.
- Use Method P2 for 8 minutes.

Note: It is normal for some proteins to remain in the gel because some high-molecular weight proteins do not transfer completely using the iBlot Gel Transfer Device, compared to semi-wet transfer apparatus.

Answer Id: E11623

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Product FAQ

I transferred my proteins using the iBlot 2 Dry Blotting System but the high-molecular weight proteins remained in the gel indicated by staining of the gel after transfer. Can you help me troubleshoot?

Answer

This could happen if an incorrect voltage Method was used or if inappropriate transfer conditions were used. Make sure that the voltage Method and run time used is correct, based on the gel type, as described on page 17 in the manual (http://tools.thermofisher.com/content/sfs/manuals/iblot2_device_man.pdf).

For mini or midi gels:
- Perform an ethanol equilibration step as described on page 35 in the manual (http://tools.thermofisher.com/content/sfs/manuals/iblot2_device_man.pdf) to improve transfer.
- Use a lower gel percentage to separate the high-molecular weight proteins.
- Increase the transfer time in 30-second increments.

For E-PAGE gels:
- Increase the transfer time in 30-second increments.
- Use Method P3 for 8 minutes.

Note: It is normal for some proteins to remain in the gel because some high-molecular weight proteins do not transfer completely using the iBlot 2 Gel Transfer Device, compared to semi-wet transfer apparatus.

Answer Id: E11640

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Product FAQ

Is it possible to do gel staining with the Power Blotter System/Power Blotter XL System?

Answer

No, the Power Blotter System/Power Blotter XL System is designed for Western transfer alone and does not support gel staining. There is no staining module available and a staining cassette is not offered.

Answer Id: E17219

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Manual / Product Insert

238570 Rapid-Chrome Kwik-Diff Staining System IFU

Version: FEB.2016
Catalog #

Product FAQ

Do you offer a staining cassette for the Power Blotter System/Power Blotter XL System?

Answer

No, the Power Blotter System/Power Blotter XL System is designed for Western transfer alone and does not support gel staining. There is no staining module available and a staining cassette is not offered.

Answer Id: E17203

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Manual / Product Insert

iBlot Dry Blotting System

Version: Rev date: 15 May 2012
Catalog #

Product FAQ

What is the difference between the Expressway Cell-Free and Expressway Lumio Cell-Free E. coli Expression Systems?

Answer

The Expressway Lumio system incorporates the benefits of the Expressway cell-free system and Lumio technology. Using the Lumio kit, your gene of interest is fused to a Lumio tag, enabling sensitive and specific in-gel detection of the Lumio -tagged fusion protein in polyacrylamide gels without the need for staining or western blotting. You can also monitor real-time synthesis of the Lumio -tagged protein using a standard fluorometer.

Answer Id: E9635

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Product FAQ

I performed stable selection but my antibiotic-resistant clones do not express my gene of interest. What could have gone wrong?

Answer

Here are possible causes and solutions:

Detection method may not be appropriate or sensitive enough:
- We recommend optimizing the detection protocol or finding more sensitive methods. If the protein is being detected by Coomassie/silver staining, we recommend doing a western blot for increased sensitivity. The presence of endogenous proteins in the lysate may obscure the protein of interest in a Coomassie/silver stain. If available, we recommend using a positive control for the western blot.
- Insufficient number of clones screened: Screen at least 20 clones.
- Inappropriate antibiotic concentration used for stable selection: Make sure the antibiotic kill curve was performed correctly. Since the potency of a given antibiotic depends upon cell type, serum, medium, and culture technique, the dose must be determined each time a stable selection is performed. Even the stable cell lines we offer may be more or less sensitive to the dose we recommend if the medium or serum is significantly different.
- Expression of gene product (even low level) may not be compatible with growth of the cell line: Use an inducible expression system.
- Negative clones may result from preferential linearization at a vector site critical for expression of the gene of interest: Linearize the vector at a site that is not critical for expression, such as within the bacterial resistance marker.

Answer Id: E9183

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Product FAQ

Can you give me an overview of the Expressway system?

Answer

- Begin by generating a DNA template, either by PCR or in a plasmid vector
- Purify the template
- Perform the synthesis reaction
- Analyze the sample via Coomassie staining, western blot, etc.

Answer Id: E9631

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Product FAQ

The power station on my G2 Fast Blotter has broken. How can I replace the station?

Answer

You can purchase the Power Blotter Station (Cat. No. PB0010) and the G2 Fast Blotter cassette is compatible with it. The new Power Blotter System is designed for Western transfer alone and does not have a staining module; therefore, the G2 Fast Blotter staining cassette cannot be used on the Power Blotter Station.

Answer Id: E17204

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Product FAQ

What are the components of the Thermo Scientific Power System?

Answer

The Thermo Scientific Power System (Cat. No. 22830) consists of the Thermo Scientific Power Station (Cat. No. 22838) with activated Staining and Blotting Software, the Thermo Scientific Power Stain Cassette (Cat. No. 22836), and the Thermo Scientific Power Blot Cassette (Cat. No. 22835). It is a combination of the Power Stainer and the Power Blotter.

Answer Id: E11228

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Manual / Product Insert

pFLD

Version: 29 March 2012
Catalog #

Manual / Product Insert

Rabbit anti-STEAP (Six Transmembrane Epithelial Antigen of the Prostate) - 2ndGen Predilute

Version: 08/08
Catalog #
  • 081385(Discontinued)

Manual / Product Insert

pIB/His A, B, and C

Version: Version C 12/29/10