Thermo Fisher Scientific Saturated Ammonium Sulfate Solution User guide

Brand: Thermo Fisher Scientific

Model: Saturated Ammonium Sulfate Solution

Type: User guide

INSTRUCTIONS

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Number Description

45216 Saturated Ammonium Sulfate Solution, 1L, sufficient solution to precipitate 1L of sample

Storage: Upon receipt store at ambient temperature.

Introduction

The Thermo Scientific™ Saturated Ammonium Sulfate Solution (4.32M) enables removal of abundant proteins from serum,

cell culture supernatant and ascites fluid. Slowly adding the Ammonium Sulfate Solution to a protein sample results in

decreased protein solubility and eventual protein precipitation as the charged ammonium sulfate ions compete with protein

for water interaction. High molecular weight proteins, such as antibodies, are among the first to precipitate. When used in

combination with antibody purification methods such as affinity chromatography, ammonium sulfate precipitation is ideal for

achieving highly purified antibody samples.

Important Product Information

• For optimal antibody recovery and minimal contaminants in the final product, add the Saturated Ammonium Sulfate

Solution slowly to the sample while stirring to avoid excessively high salt concentrations at the site of addition.

• The precipitated antibody must be extensively dialyzed or desalted before most downstream applications.

• If the bottle is left open and evaporation occurs, the ammonium sulfate may precipitate; however, this occurrence does

not affect functionality of the solution. Avoid transferring ammonium sulfate crystals to the sample. If desired, filter the

solution to remove crystals.

Procedure for Antibody Precipitation1

1. Place the sample in a beaker or flask that can contain twice the sample’s volume. Place the container on a stir plate and

add a stir bar.

2. While stirring, slowly add a volume of the Saturated Ammonium Sulfate Solution equal to the sample volume.

Note: For optimal antibody recovery and minimal contaminants in the final product, add the Saturated Ammonium

Sulfate Solution slowly to the sample while stirring to avoid excessively high salt concentrations at the site of addition.

3. Allow sample to precipitate for 30 minutes at room temperature or 6 hours to overnight at 4°C.

4. Centrifuge sample at 3000 × g for 20 minutes.

5. Discard supernatant and dissolve the precipitate in a desired buffer. Use a volume of buffer from 0.1 to 1.0 times the

original sample volume.

6. Prepare dialysis buffer at 300-500 times the volume of the sample and perform dialysis as follows:

 Dialyze for 2 hours at room temperature

 Change the dialysis buffer and dialyze for another 2 hours.

 Change the dialysis buffer and dialyze overnight at 4°C.

Note: For samples < 4mL, desalting columns (e.g., Thermo Scientific™ Zeba™ Spin Desalting Columns) can be

substituted for dialysis.

1757.2

45216

Saturated Ammonium Sulfate Solution

Pierce Biotechnology

PO Box 117

(815) 968-0747

www.thermoscientific.com/pierce

3747 N. Meridian Road

Rockford, lL 61105 USA

(815) 968-7316 fax

Information Available from Our Website

• Tech Tip #43: Protein stability and storage

• Tech Tip #6: Extinction coefficient guide

• Tech Tip #20: Dialysis: an overview

Related Thermo Scientific Products

89882 Zeba Spin Desalting Columns, 0.5mL, 25 columns, for 30-130µL samples

89889 Zeba Spin Desalting Columns, 2mL, 5 columns, for 200-700µL samples

89891 Zeba Spin Desalting Columns, 5mL, 5 columns, for 500-2000µL samples

89893 Zeba Spin Desalting Columns, 10mL, 5 columns, for 700-4000µL samples

68100 SnakeSkin™ Dialysis Tubing, 10K MWCO, 22mm dry I.D. × 35 feet

68011 SnakeSkin Dialysis Tubing Clips, 6 each

66382 Slide-A-Lyzer™ Dialysis Cassette Kit, 10 dialysis cassettes, each appropriate for 0.5-3.0mL samples

66807 Slide-A-Lyzer Dialysis Cassette Kit, 10 dialysis cassettes, each appropriate for 3-12mL samples

66830 Slide-A-Lyzer Dialysis Cassette Kit, 10 dialysis cassettes, each appropriate for 12-30mL samples

66493 Slide-A-Lyzer Syringe (20mL) and 18-Gauge Needles, 10 each

66529 Slide-A-Lyzer Concentrating Solution, 500mL

25200-25244 Precise™ Protein Gels (see catalog or web site for a complete listing)

Cited Reference

1. Harlow, E. and Lane, D. (1988). Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory; New York: p. 298-9.

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