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Manual / Product Insert

Product Info Sheet: iBright Prestained Protein Ladder

Version: MAN0016264 Rev. A.0 (01Mar2017)

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

I used one of your pre-stained protein standards for a western transfer and I noticed that the intensity of the band faded from the membrane during the transfer process. Why is this?

Answer

The fading is most likely due to detergent in the western blocking/washing solutions that can remove some of the proteins from the membrane. The dye itself will not wash off of the proteins because it is covalently bound. We have found that smaller pore size membranes retain the proteins better during blocking and wash procedures, and hence recommend use of 0.2 µm instead of 0.45 µm membranes for best resolution and protein retention. After transfer, it is a good idea to circle the pre-stained bands with a pencil on the membrane, so band positions can be identified after blocking and processing.

Answer Id: E11760

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Product FAQ

Are any bovine-derived proteins present in PageRuler and Spectra Prestained Protein Ladders?

Answer

All PageRuler and Spectra ladders are made out of recombinant prokaryotic proteins and do not contain any animal-derived proteins.

Answer Id: E8794

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Manual / Product Insert

User Guide: Spectra Multicolor Low Range Protein Ladder

Version: B.0
Catalog #

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

What are the storage conditions and shelf life for the BenchMark Prestained Protein Ladder?

Answer

We recommend storing the BenchMark Prestained Protein Ladder at -20 degrees C. It is stable for 1 year when properly stored.

Answer Id: E11679

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Manual / Product Insert

Product Information Sheet: iBright Prestained Protein Ladder

Version: MAN0016682 Rev. E.0

Product FAQ

Can the BenchMark His-tagged Protein Standard be detected using anti-HisG antibody?

Answer

The BenchMark His-tagged Protein Standard cannot be detected using anti-HisG antibody. It can be detected using Anti-His(C-term) Antibody, Cat. No. R930-25.

Answer Id: E11717

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Product FAQ

I used one of your protein standards and am seeing some extra bands in the lane. Can you offer some suggestions?

Answer

- While loading, take care to make sure that there is no cross-contamination from adjacent sample lanes.
- Make sure that the correct amount of standard is loaded per lane. Loading too much protein can result in extra bands and this is a problem especially with silver-stained gels.
- Improper storage of the standard or repeated freeze/thawing can result in protein degradation.

Answer Id: E11756

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Product FAQ

I used your MagicMark XP Western Protein Standard and obtained a very weak or almost no signal after the western detection. Can you please help me troubleshoot?

Answer

Here are some suggestions:

- Verify that the detection reagents are working well. Optimize the antibody concentration to obtain best results.
- Make sure that the amount of standard loaded on the gel is correct.
- Optimize the transfer conditions (current, voltage, transfer time).
- Enzyme-conjugated primary antibodies may not bind efficiently with the proteins in the MagicMark XP Western Protein Standard. We recommend using unconjugated primary antibody, followed by the addition of enzyme-conjugated secondary antibody.
Note: The Anti-myc-AP/HRP and Anti-V5-AP/HRP antibodies do not bind to MagicMark XP proteins.

Answer Id: E11761

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Product FAQ

What are the concentrations of the individual proteins in the Thermo Scientific protein ladders? Is it possible to use the ladders as a standard for protein quantification?

Answer

Thermo Scientific ladders are not designed for protein quantification. For quantification, we would recommend to use a protein of known concentration as a reference.

Answer Id: E8795

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Product FAQ

I used one of your Thermo Scientific Spectra prestained protein ladders for a western transfer and got very poor transfer onto the membrane. What possibly went wrong?

Answer

Here are possible causes and solutions:

- Not enough volume of ladder loaded on the gel: Load an appropriate volume of the ladder onto the gel. Here are our recommendations:
--- Mini-gel: 5 µL per well (0.75-1.0 mm thick) or 10 µL per well (1.5 mm thick)
--- Large gel: 10 µL per well (0.75-1.0 mm thick) or 20 µL per well (1.5 mm thick)
- Incomplete or poor transfer: Optimize transfer conditions

Answer Id: E11765

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