Streptavidin Agarose Beads

Overview

Description

Streptavidin Agarose Beads is useful for the precipitation of biotinylated proteins. Recombinant streptavidin is immobilized via covalent binding of primary amino groups to N-hydroxysuccinimide (NHS)-activated agarose beads.

Tested applications

For coupling of proteins, antibodies, peptides and other molecules. Optimal amount should be determined by the end user.

Product Picture

Properties

Form

Beads in suspension (50% slurry), supplied in phosphate buffered saline (PBS) pH 7.4, with 0.02% sodium azide as preservatives.

Storage instruction

Store at 4°C. This product is stable for 12 months. Do not freeze. 

Beads Diameter

90 µm

Binding Capacity

>100nmol Free biotin

200μg Biotinylated antibody

(/mL beads slurry)

Applications

Application Image

Protocols

Additional information

 

• Streptavidin Agarose Beads should be resuspended well before used. 
•  Beads cannot be reused.

 

Procedure for binding biotinylated target directly.

 

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

 

1. Recommended coupling/washing buffers

protein/antibody samples: PBS buffer, pH7.4, nucleic acids samples: TES buffer

 

2. Elution buffer (only for biotinylated target purification): 8M Guanidine hydrochloride, pH1.5

 

B. Wash Streptavidin agarose beads

Calculate the amount of beads required based on their binding capacity (see the above Binding Capacity), and transfer the beads to a new tube.
 

1. Resuspend the beads in the vial (i.e. tilt and rotate for 5 min).

2. Transfer the desired volume of beads to a tube.

3. Add an equal volume of Washing buffer, or at least 1 mL, and mix (keep on a roller for at least 5 min).

4. Centrifuge the tube at 4000 g for 1 min and discard the supernatant.

5. Resuspend the washed beads in the same volume of washing buffer as the initial volume of beads taken from the vial (step 2).

 

C. Binding protein/antibody

1. Incubate the beads and biotinylated protein/antibody in PBS buffer (pH 7.4) for 30 min at room temperature using gentle rotation.

2. Centrifuge the protein/antibody-coated beads at 4000 g for 2–3 min and discard the supernatant.

3. Wash the coated beads 4–5 times in PBS containing 0.1% BSA.

4. Resuspend to the desired concentration for your application.

 

NOTE:

Elution of the biotinylated proteins that are bound to the Streptavidin Beads requires harsh conditions, such as boiling for 5 minutes with 8M guanidine•HCl, pH 1.5. Protein may leach from the beads with such conditions after centrifugation.