Product FAQ

What are the apparent molecular weights of the proteins in the BenchMark Fluorescent Protein Standard?

Answer

The BenchMark Fluorescent Protein Standard is designed for visualization of molecular weight ranges of proteins labeled with Lumio Green Detection kit (Cat. No. LC6090) or other fluorescent-conjugated proteins. The standard consists of a set of wide molecular weight range proteins conjugated to a fluorescent dye, Alexa Fluor 488. The standard can also be detected with Coomassie staining. It is suitable for use with NuPAGE or Tris-Glycine gels. Here are the apparent molecular weights of the proteins in the BenchMark Fluorescent Protein Standard in the different gels:

- Band 1: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 155 kda; 4-20% Tris-Glycine gel - 155 kDa
- Band 2: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 98 kDa; 4-20% Tris-Glycine gel - 100 kDa
- Band 3: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 63 kDa; 4-20% Tris-Glycine gel - 65 kDa
- Band 4: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 40 kDa; 4-20% Tris-Glycine gel - 41 kDa
- Band 5: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 32 kDa; 4-20% Tris-Glycine gel - 33 kDa
- Band 6: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 21 kDa; 4-20% Tris-Glycine gel - 23 kDa
- Band 7: NuPAGE 4-12% Bis-Tris gel with MES/MOPS buffer - 11 kDa; 4-20% Tris-Glycine gel - 12 kDa

Answer Id: E11721

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Product FAQ

What are the storage conditions and shelf life for the BenchMark Fluorescent Protein Standard?

Answer

We recommend storing the BenchMark Fluorescent Protein Standard in the dark at -20 degrees C and in aliquots to avoid repeated freezing and thawing. It is stable for 6 months when properly stored.

Answer Id: E11720

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Manual / Product Insert

BenchMark Fluorescent Protein Standard

Version: 17 January 2012
Catalog #

Product FAQ

What is the identity of each protein band in the BenchMark Fluorescent Protein Standard?

Answer

The identity of each protein band in the BenchMark Fluorescent Protein Standard is proprietary.

Answer Id: E11724

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Product FAQ

Can I use the BenchMark Fluorescent Protein Standard for protein quantitation?

Answer

The concentration of each protein in the standard is proprietary. We do not recommend using the standard for quantitation purpose.

Answer Id: E11723

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Product FAQ

What are the maximum excitation and emission wavelengths for the Alexa Fluor 488 dye that is bound to the proteins in the BenchMark Fluorescent Protein Standard?

Answer

The maximum excitation of the dye is at 493 nm and the maximum emission is at 516 nm.

Answer Id: E11722

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Product FAQ

Can you suggest a suitable protein standard to visualize my Lumio fusion proteins?

Answer

The BenchMark Fluorescent Protein Standard allows direct visualization of molecular weight ranges of Lumio fusion proteins on an SDS-PAGE gel. The proteins in the standard are easily detected using a UV transilluminator or a laser-based scanner at the same excitation and emission wavelengths as your Lumio fusion protein.

Answer Id: E11190

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Product FAQ

Which protein standards do you recommend using with E-PAGE 48 and E-PAGE 96 gels for electrophoresis, Western blotting and fluorescent detection?

Answer

Electrophoresis:
For E-PAGE 48 8% gels, use SeeBlue Plus 2 Pre-Stained Standard, Cat. No. LC5925.
For E-PAGE 96 6% gels, use E-PAGE SeeBlue Pre-Stained Standard, Cat. No. LC5700.

Western Blotting:
For E-PAGE 48 8% gels, use MagicMark XP Western Protein Standard, Cat. No. LC5602.
For E-PAGE 96 6% gels, use E-PAGE MagicMark Unstained Protein Standard, Cat. No. LC5701.

Fluorescence detection:
For E-PAGE 48 8% gels, use BenchMark Fluorescent Protein Standard, Cat. No. LC5928.
For E-PAGE 96 6% gels, use BenchMark Fluorescent Protein Standard, Cat. No. LC5928.

Answer Id: E10771

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Product FAQ

What protein markers should be used for E-PAGE gels?

Answer

The following protein MW standards are recommended for E-PAGE 96 gels.

Cat. No. LC5700 E-PAGE SeeBlue Pre-Stained Standard for general electrophoresis
Cat. No. LC5701 E-PAGE MagicMark Unstained Protein Standard for Western blotting
Cat. No. LC5928 BenchMark Fluorescent Protein Standard for fluorescent detection

The following protein MW standards are recommended for E-PAGE 48 gels.

Cat. No. LC5925 SeeBlue Plus 2 Pre-Stained Standard for general electrophoresis
Cat. No. LC5602 MagicMark XP Western Protein Standard for Western blotting
Cat. No. LC5928 BenchMark Fluorescent Protein Standard for fluorescent detection

Answer Id: E4857

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Product FAQ

I used the Lumio Green Detection Kit and am getting a very weak signal. Why is this?

Answer

Here are possible causes and solutions:

- Improper labeling. Make sure that the labeling protocol is correctly followed to obtain the best results. Make sure you have added the Lumio Green Detection Reagent to the samples prior to electrophoresis. Limit exposure of the Lumio Gel Sample Buffer (4X) to air. Always return the Lumio Green Reagent and Lumio Enhancer to ?20 degrees C immediately after use to preserve the activity of buffers.
- Low protein load or low expression level. Check total protein loaded on the gel by staining the gel with a total protein stain as described in the manual. Load at least 1 pmole of the Lumio fusion protein. Make sure the Lumio tag is in frame and the protein is expressed properly. A positive control is supplied with the Lumio vectors to verify the expression protocol.
- The gel is exposed to UV light for a long time. The fluorescent dye of the Lumio Green Reagent is sensitive to photobleaching, so avoid exposing the gel to UV light for a long time.
- The gel is not visualized immediately or imaged properly. Be sure to visualize the gel after removing the gel from the cassette and view the gel immediately after electrophoresis. Use a UV transilluminator or a laser-based scanner using appropriate filters as described in the manual. Tip: If you have run BenchMark Fluorescent Protein Standard on the same gel and can view the standard bands on the gel, then you are imaging the gel properly.

Answer Id: E11288

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Citations & References

Protistan grazing analysis by flow cytometry using prey labeled by in vivo expression of fluorescent proteins.

  • Authors: Fu Y, O'Kelly C, Sieracki M, Distel DL
  • Journal: Appl Environ Microbiol (2003) 69:6848-6855
  • PubMed ID: 14602649
Catalog #

Manual / Product Insert

Lumio Green Detection Kit

Version: 8 March 2012
Catalog #

Product Literature

Lumio™ Technology

Manual / Product Insert

Expressway HTP Cell-Free E. coli Expression Kit

Version: Version C 11/20/10

Product Literature

Protein transfer technical handbook