Krypton™ Fluorescent Protein Stain - FAQs

查看更多产品信息 Krypton™ Fluorescent Protein Stain - FAQs (46630)

14 个常见问题解答

If the Krypton Protein Stain is cloudy, can I still use it?

Yes. The solution will be cloudy immediately upon dilution to 1X, especially if it is cold. The cloudiness should dissipate with mixing.

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

What is the storage temperature for the Krypton Protein Stain and how stable is it?

Store Krypton Protein Stain at 4 degrees C; although it is stable enough for ambient shipping. The 10X solution is under warranty for one year from the date of shipment if handled and stored properly. The Working Solution (1X) is stable at 4 degrees C for 1-2 weeks.

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How is the Krypton Protein Stain's signal captured?

Filter sets designed for Cy3 will work with the Krypton Protein Stain. Thermo Scientific scientists have used GE Healthcare Typhoon 9410 Variable Mode Imager (532 nm laser excitation; 580 BP 30 emission filter), Kodak Imager (CCD; 535 nm excitation and 600 nm emission filters), and visible light box. Sensitivity is better with lasers than with transilluminators.

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

Is Krypton Protein Stain compatible with 2-D electrophoresis and mass spectrometry?

Yes. HeLa cell and rat heart tissue lysates separated by 2D electrophoresis and stained with the Krypton Protein Stain provided excellent sensitivity while not over-saturating highly concentrated spots. MALDI tryptic peptide mass fingerprinting of BSA and myoglobin resulted in characteristic profiles. The proteins were destained and processed with the Thermo Scientific In-Gel Tryptic Digestion Kit (Cat. No. 89871). The lack of peak shifts indicates that the dye was not bound to the proteins after destaining.

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

Does Krypton Protein Stain label nucleic acids?

No.

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Has the Krypton Protein Stain been compared to other stains?

Yes. Bio-Rad Flamingo, GE Healthcare Deep Purple and Invitrogen SYPRO Ruby Stains, all had problems not observed with Krypton Protein Stain. For example, Flamingo Stain saturates and requires a full five hours for staining, Deep Purple Stain produces fuzzy spots on 2D gels and SYPRO Ruby Stain is expensive and the least sensitive of the four.

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

How photostable is Krypton Protein Stain?

Krypton Protein Stain has good photostability. Typically, only an 8% decrease in signal occurs after 16 sequential scans.

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Does the Krypton Protein Stain produce a linear response for quantitative analysis?

Yes. The Krypton Protein Stain produces a linear quantitative range of three-to-four orders of magnitude.

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

Why are so many proteins easily stained with the Krypton Protein Stain?

Ionic interactions cause the stain to bind basic amino acid residues, and hydrophobic interactions allow it to bind to alkyl chains. The dual-binding action makes the Krypton Protein Stain a universal stain for protein staining.

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How long does it take to stain gels using the Krypton Protein Stain?

The standard protocol takes only 2.5 hours to complete. A rapid, 30 minute protocol is also available that is sensitive down to 2 ng.

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

What are the excitation/emission maxima and the extinction coefficient for the Krypton Protein Stain?

The excitation/emission maxima are 520 nm/580 nm with an extinction coefficient of 105,000 per mol cm.

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What gel types have been used with the Krypton Protein Stain?

Krypton Protein Stain has been used with Thermo Scientific Precise Protein Gels (Tris-HEPES), Invitrogen Invitrogen Gels (Tris-glycine) and NuPAGE (Bis-Tris) and Bio-Rad Criterion Gels (Tris-HCl). Krypton Protein Stain is also compatible with homemade gels, native gels and 2D gels. Sensitivity may vary depending on gel type.

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

What is the sensitivity of Krypton Protein Stain?

The stain is sensitive down to 0.25 ng using the standard protocol, or down to 2 ng using the rapid 30 minute protocol. A broad range of proteins have been tested with molecular weights from 14.4 to 200 K, including glycoproteins and phosphoproteins. Some of the proteins tested are as follows: myosin, beta-galactosidase, phosphorylase B, bovine serum albumin, ovalbumin, carbonic anhydrase, soybean trypsin inhibitor, lysozyme, aprotinin, horseradish peroxidase, myokinase, avidin, glucose oxidase, a2-macroglobulin, a1-acid glycoprotein, and beta-casein. (HeLa cell and rat heart tissue lysates were tested with 2D applications).

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

Can I use Krypton Fluorescent Protein Stain (Cat. No. 46630) with UV light?

We don't recommend using Krypton Fluorescent Protein Stain with UV light because the sensitivity of the stain decreases by four-five fold in UV light. Instead, we recommend using SYPRO Ruby Protein Gel Stain (Cat. Nos. S12000, S12001, S21900).

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