Pierce™ 6xHis Protein Tag Stain Reagent Set
Pierce™ 6xHis Protein Tag Stain Reagent Set
Thermo Scientific™

Pierce™ 6xHis Protein Tag Stain Reagent Set

The Thermo Scientific Pierce 6xHis Protein Tag Stain Reagent Set is a fluorescent stain for preferential detection of histidine-tagged (His-tagged)Read more
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Catalog number 24570
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448.00
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Price (USD)
448.00
Each
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The Thermo Scientific Pierce 6xHis Protein Tag Stain Reagent Set is a fluorescent stain for preferential detection of histidine-tagged (His-tagged) recombinant fusion proteins in protein polyacrylamide gels.

The unique fluorescent stain specifically detects histidine-tagged fusion proteins that have been electrophoresed in polyacrylamide gels (e.g., SDS-PAGE). With this stain, expression of His-tagged recombinant proteins can be assessed directly in-gel, eliminating membrane transfer and Western blotting steps that are typically used to confirm purity and yield of tagged proteins. Although the lower limit of detection depends on which UV lamp (300nm) and detection equipment (CCD camera best) are used, the stain is convenient for in-process assessment of His-tagged protein production and is compatible with subsequent staining with coomassie dye and other total protein stains.

Features of the 6xHis Protein Tag Stain Reagent Set:

• Two- to three-times faster than Western blotting—get results sooner and save hours of valuable research time
• Detects directly on the gel—can eliminate the need for a membrane transfer and Western blot step for the detection of at least 0.2 μg of 6xHis-tagged protein
• Ready-to-use, two-reagent formula—no mixing, no diluting and no fuss; guarantees a simple-to-perform, mistake-free detection protocol
• Specific fluorescent detection of 6xHis-tagged proteins—see only what you want to see (CCD camera recommended for low abundance proteins; UV-transilluminator for abundant proteins)
• Compatible with double-staining—follow His-tagged protein staining with Thermo Scientific GelCode Blue Stain Reagent for a total protein profile determination

Includes:
• Stain and developer solutions (500 mL each)

Requires:
• UV lamp source (300nm) and CCD camera
For Research Use Only. Not for use in diagnostic procedures.
Specifications
DescriptionPierce 6xHis Protein Tag Stain Reagent Set
Detection LocationIn-Gel Detection
Detection MethodFluorescence
Label or DyeProprietary Mix
Product TypeHis Tag Protein Stain Set
Product LinePierce™
Quantity1 L
Target MoleculeProteins (His-Tagged)
Unit SizeEach
Contents & Storage
Store components at room temperature. Product is shipped at ambient temperature

Frequently asked questions (FAQs)

I used the Thermo Scientific 6xHis Protein Tag Stain Reagent Set to detect my 6xHis-tagged protein but am also detecting non-tagged proteins. Why is this?

This is likely due to weak cross-reaction staining of proteins containing histidine clusters. Here are our recommendations:

- Wash gel for additional time in water (step 5.)
- Slightly decrease staining time (step 2.)
- Adjust exposure time and other settings to minimize weak, non-specific staining.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

With the Thermo Scientific 6xHis Protein Tag Stain Reagent Set, I was able to detect the positive control but not the experimental protein. What happened?

Here are possible causes and solutions:

- Experimental protein not expressed at sufficient levels in the lysate being tested. Load more lysate per lane or otherwise check that the target protein is expressed at all.
- Experimental recombinant protein is not tagged with 6xHis. Check for presence of tag by an independent method (e.g., detection or purification by nickel-chelate chemistry.
- 6xHis tag on experimental protein is blocked by interfering substances in sample. Verify that nickel and other 6xHis-binding reagents were not brought forward from a previous step and use only high-quality water.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I used the Thermo Scientific 6xHis Protein Tag Stain Reagent Set but was not able to detect any bands for either the positive control or experimental protein. What possibly went wrong?

Here are possible causes and solutions:

- Poor quality or insufficient exposure to appropriate UV-light source. If possible, use a CCD camera for detection; ensure that UV lamp delivers the appropriate wavelength for excitation (280-310 nm).
- Experimental protein is poorly expressed (insufficient loading). Insufficient protein was electrophoresed per lane for the detection method used.
- Insufficient washing; residual SDS in gel prevents binding of stain. Wash gel for 3 × 20 minutes in ultrapure water and restain .
- Experimental protein is small (less than 20kDa) and diffused from gel during washing step. Fix the gel 50% methanol:7% acetic acid for 15 minutes before performing the water wash.
- Poor diffusion of stain into gel. Increase staining time to 10 minutes (step 2); this may be repeated on the same gel.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

With the Thermo Scientific 6xHis Protein Tag Stain Reagent Set, how long will the fluorescent signal remain stable after the gel is stained'?

The fluorescent signal is stable for several hours in gels stored in water. Signal may be detectable, if somewhat attenuated, after overnight storage.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Is there a fixation step in the Thermo Scientific 6xHis Protein Tag Stain Reagent Set staining procedure?

There is no fixation step in the Thermo Scientific 6xHis Protein Tag Stain Reagent Set staining procedure. Hence, staining with this kit does not inhibit subsequent total protein staining with general protein stains, or electrophoretic transfer to membrane. Note: Bis-Tris gels run in MOPS or MES buffer may require fixing in 50% methanol: 7% acetic acid for 15 minutes before performing the stain procedure. After electrophoresis, fix the gel and then proceed with Step 1 of the procedure.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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