Bio-Rad Silver Stains User Manual

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Life Science
Group

SIG 121393 Printed in USA

Bio-Rad
Laboratories

Bio-Rad Silver Stain

Catalog Numbers

161-0443
161-0444
161-0445
161-0447

For Technical Service

Call Your Local Bio-Rad Office or

in the U.S. Call 1-800-4BIORAD

(1-800-424-6723)

LIT34 Rev B

Table of Contents

Section 1 Introduction...............................................................1

Section 2 Silver Stain Kit Components ...................................1

Section 3 Important Procedural Information.........................1

3.1 Water Purity.........................................................................1

3.2 Gel Handling .......................................................................2

3.3 Temperature.........................................................................2

3.4 Convenient Stopping Point..................................................2

3.5 Band Development..............................................................2

3.6 Destaining............................................................................2

3.7 Storage of Stained Gels.......................................................3

3.8 Photographing Gels.............................................................3

Section 4 Reagent Preparation.................................................4

Section 5 Protocols ....................................................................5

Section 6 Additional Applications............................................7

6.1 Nucleic Acid Staining..........................................................7

6.2 Silver Staining after Coomassie Blue R-250 Staining.........7

6.3 Silver Staining Electrofocusing Slab Gels...........................7

Section 7 Troubleshooting Guide.............................................7

Section 8 Ordering Information...............................................9

Section 9 References................................................................10

9.1 General ..............................................................................10

9.2 Proteins..............................................................................10

9.3 Nucleic Acids ....................................................................11

9.4 Isoelectric Focusing...........................................................11

Section 1
Introduction

Silver staining is a highly sensitive method for detecting proteins and nucleic
acids in polyacrylamide slab gels. The Bio-Rad Silver Stain, derived from the
method of Merril,1 is 10-50 fold more sensitive than Coomassie brilliant blue
R-250 for proteins (detection is ~0.1 ng/mm2) and 2-5 fold more sensitive than ethid­ium bromide for single and double stranded DNA and RN A. Approximately 24 full
size gels or 48 mini gels can be stained with one kit.

Section 2
Silver Stain Kit Components

Oxidizer Concentrate - A 10-fold stock solution. Contains potassium dichro­mate (CAS 7778-50-9) and nitric acid. Store at 4 ˚C.

Warning: Strong oxidizer - avoid contact with reducing agents.

Irritant - avoid contact with skin and eyes. Wear gloves and eye pro-

tection.

Silver Reagent Concentrate - a 10-fold stock solution. Contains silver nitrate
(CAS 7761-88-8). This solution is heat sensitive. Store at 4 ˚C.
Warning: Poisonous - caustic to eyes, skin, and mucous membranes. Wear

gloves and eye protection.

Developer - Four packages of dry chemical blend. Contains sodium carbonate and
paraformaldehyde (CAS 30525-89-4). Store at 4 ˚C.

Warning: Poisonous

Irritant - vapor and dust irritates eyes, mucous membranes, and skin.

Wear gloves, lab coat, and eye protection. Fume hood recommended.

Section 3
Important Procedural Information

Read the entire protocol, procedural information, and the Troubleshooting
Guide before beginning.

3.1 Water Purity

Deionized water of less than 1 µmho conductivity is recommended for all phas­es of the procedure. Contaminants such as chloride ions will precipitate silver
ions causing reduced sensitivity and increased background. Water suspected of
containing interfering ions can be thoroughly deionized by passage over a column

®

of AG

501-X8(D) mixed bed ion exchange resin (catalog number 142-6452).

1