2-D Electrophoresis Workflow
Oriole™ Fluorescent
Gel Stain
Instruction Manual
Catalog #
161-0495, 1x solution, 200 ml
161-0496, 1x solution, 1 L
161-0497, kit for 5 L
For technical suppor t call yo ur local Bio-Rad offi ce.
In the U.S. ca ll 1-800-424-6723.
Table of Contents
Section 1: Introduction an d General Informat ion 1
1.1 Introduction 1
1.2 Product D escription 2
1.3 Storage 2
1.4 Materials and Equipment Required
but Not Supplied 3
1.5 Reagents Required but Not Supplied 3
1.6 Safety Considerations 4
1.7 Disposal Considerations 4
1.8 Fluorescence Characteristics 4
Section 2: Instru ctions 6
2.1 General Considerations 6
2.2 Stain Solution Preparation 8
2.3 Gel Staining 9
2.4 Gel Imaging 11
Section 3: Technical Information 14
3.1 Sensitivity of Staining — 1-D Gels 14
3.2 Compatibilit y with Mass Spectrometr y 15
3.3 Protein-to-Protein Variability 15
3.4 Dynamic Range 16
3.5 2-D Gel Staining 17
Section 4: Troubleshooting 18
Section 5: Product Information 21
™
5.1 Oriole
Fluorescent Gel Stain 21
5.2 Related Products 21
Section 1
Introduction and General Information
1.1. Introduction
™
fluorescent gel stain is an easy to use, rapid, and
Oriole
sensitive stain for visualization and quantitation of proteins
separated by SDS-PAGE. The product is available in
three configurations. The 200 ml and 1 L sizes are
provided ready to use. The product is also available as a
kit containing components to make 5 L of ready to use
staining solution.
The staining procedure is a simple one-step protocol
that can be completed in as little as 9 0 minutes.
Gels stained with Oriole fluore scent gel stain may
be visualized with a variety of dif ferent UV-based
fluorescence imaging systems.
Oriole fluorescent gel stain gives exceptional sensitivity
and dynamic range (see pages 14–16) and is compatible
with subsequent analysis by enzymatic digestion and
mass spectrometry. It is thus particularly well suited to
proteomics applications.
1
1.2. Product Description
Oriole fluorescent gel stain comes in three package
configurations.
The 200 ml size — fully diluted and ready to use;
provide s enough stain for four mini format
Mini-PROTEAN
Criterion
®
gels (~8.6 x 6.8 cm), or two midi format
™
gels (13.3 x 8.7 cm).
The 1 L size — fully diluted and ready to use; provides
enough stain for 20 Mini-PROTEAN gels (~8.6 x 6.8
cm), ten Criter ion gels (13.3 x 8.7 cm), four large format
PROTE AN
®
II gels (16 x 16 cm or 16 x 20 cm), or two large
format PROTEAN Plus gels (25 x 20.5 cm).
The 5 L kit — contains concentrated compone nts to
prepare 5 L of staining solution and can be diluted to
1x according to demand.
1.3. Storage
The product is sta ble for at least 18 months from the date
of manufacture or until the expiration date on the label
when stored at 24°C or below. Consult the expiration date
2
before using. Avoid prolonged exposure to temperatures
greater than 37°C and protect from light.
1.4. Materials a nd Equipment Required but
Not Sup plied
n
Staining containers — Any glass or plastic tray capable
of holding the recommended volume of solution may
be used
n
Imaging equipment — Gels are best imaged using a
UV-based fluorescence imager capable of excitation
near 270 nm and detection near 604 nm such as the
Molecular Imager
ChemiDoc
®
Gel Doc™ XR+, Molecular Imager®
™
XRS+, VersaDoc™ MP 4000, ExQuest™
spot cutter, and VersaDoc MP 500 0 systems. For a
more complete list of compatible imaging systems,
see pages 12–13
n
L aboratory shaker or rocker
n
Powder-free latex, vinyl, or nitrile gloves
1.5. Reag ents Required but Not Supplied
Methanol, reage nt grade (for 5 L kit only)
3
1.6. Safet y Consideration s
Oriole fluorescent gel stain is a dilute solution of a
fluorescent dye. T he working solution is flammable and
should be handled in a manner that prevents exposure to
open flame or sparks. The complete proper ties of the dye
component have not been investigated. Eye protection
and gloves should be worn and general laboratory safety
precautions followed while handling both the diluted and
undiluted product.
1.7. Disposal Considerations
Laws governing the disposal of laboratory chemicals
vary by re gion. Consult the MSDS (available online at
www.bio-rad.com) and check local laws for proper
disposal guidelines.
1.8. Fluorescence Characteristics
Oriole fluorescent gel stain has a fluorescence excitation
maximum of 270 nm and a fluorescence emission
maximum of 604 nm.
4
120
100
80
60
40
20
Fluore scence, norm alized
0
200 300 40 0 500 600 700 800
Fig. 1. Fluo resce nce exci tatio n and emissio n spec tra of
Oriol e stai n. Orio le stai n has its exc itatio n maxi mum at 270 nm
and emi ssion m axim um at 604 n m, making it compatible with
UV-base d image rs. —, Excitation sp ectru m;
Wavelen gth, nm
....
, emiss ion spe ctrum.
5
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