Thermo Scientific™ Krypton™ Fluorescent Protein Stain

Catalog No.	Quantity
46-630	500 mL

Catalog No. 46-630 Supplier Thermo Scientific™ Supplier No. 46630

Description

Stain proteins in polyacrylamide gels or transfer membranes for fluorescent detection with popular fluorescence imagers.

Thermo Scientific™ Pierce Krypton Fluorescent Protein Stain enables sensitive fluorescent visualization (520nm excitation; 580nm emission) of proteins separated by polyacrylamide gel electrophoresis or on transfer membranes using popular commercial fluorescence imagers.

This fluorescent protein gel stain is for bright yellow-orange fluorescent detection of proteins separated by 1D or 2D polyacrylamide gel electrophoresis (SDS-PAGE). Krypton Stain uses a protein-binding dye with fluorescence properties (Ex/Em = 520/580nm) suitable for visualization with a variety of standard laser-based and filter-based fluorescence imagers and documentation systems. Proteins are detectable at concentrations greater than 0.25ng per band in a typical mini-gel lane (greater than 2ng for the rapid staining protocol). Krypton Stain provides high signal intensity, exhibits minimal protein-to-protein variation, has an excellent quantitative dynamic range and is compatible with subsequent destaining and analysis by mass spectrometry.

Highlights:

• Intense fluorescent stain – excitation and emission maxima of 520nm and 580nm, respectively (compatible with 532nm excitation light sources and 580 to 600nm emission filters)

• Instrument-compatible – effective with popular laser-based and filter-based fluorescence imagers

• Broad dynamic range – linear quantitative range spans three to four orders of magnitude

• Sensitive – detect as little as 0.25ng protein with the standard protocol (approx. 2.5 hours); at least as sensitive as fluorescent stains from other suppliers

• Fast – standard (2.5 hours) and rapid (30 minutes) protocols are significantly faster than procedures with other popular fluorescent stains

• Versatile – compatible with proteomics workflows involving downstream mass spectrometry

• Compact – supplied as a 10X solution to save storage space; just dilute with water before use

Specifications

• Content And Storage: Upon receipt store at 4°C.
• Detection Location: In-Gel Detection
• Detection Method: Fluorescence
• Label or Dye: Proprietary Mix
• Quantity: 500 mL
• Target Molecule: Protein
• Product Line: Krypton
• Product Type: Protein Stain

Frequently Asked Questions (FAQs)

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.

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.

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.

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.

Does Krypton Protein Stain label nucleic acids?

No.

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.

How photostable is Krypton Protein Stain?

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

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.

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.

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.

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.

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.

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).

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).

For Research Use Only. Not for use in diagnostic procedures.