Protein A Magnetic Beads

Easy to use, high-binding capacity, non-adherent beads.

Support Characteristics: Paramagnetic, spherical, 6 % cross-linked agarose.

Ligand: Recombinant Protein A.

Particle Size: 75 – 150 μm.

Binding Capacity: Generally >25 mg human IgG/ml wet beads.

Working Temperature: Room temperature.

Storage Solution: PBS with 0.02% Sodium Azide.

Storage Temperature: 4 – 8 °C.

Applications:

Useful for immunoprecipitation and enrichment of IgG antibodies.

High affinity for Fc region of IgG antibodies from a variety of species.

Protein A binds to most human and mouse IgG subclasses (e.g., human IgG1, IgG2, IgG4; mouse IgG1, IgG2a, IgG2b, IgG3).

It also binds to total IgG from cow, guinea pig, hamster, horse, pig, and rabbit. Protein A has little affinity to chicken, goat, rat and sheep.

This product is manufactured by BioVision, an Abcam company and was previously called 6507 Protein A Magnetic Beads. 6507-1 is the same size as the 1 ml size of ab214286.

Description:

Protein A Magnetic Beads are prepared by covalently coupling Recombinant Protein A to 6% crosslinked magnetically beaded agarose. The coupling technique is optimized to give a high binding capacity for IgG. The capacity of IgG binding is generally greater than 25 mg of human IgG per ml of wet gel.

SUGGESTED PROTOCOL:

Prepare the antibody solution by diluting the required amount of antibody in binding buffer before running the protocol.

1. Magnetic Bead Preparation (perform three times)

a. Dispense the required amount of magnetic beads into a 1.5 ml microfuge tube.

b. Place the tube in the magnetic rack and remove the storage solution.

c. Add 500 μl binding buffer.

d. Resuspend the beads.

e. Remove the liquid

2. Antibody Capture

a. Immediately add the antibody solution.

b. Resuspend and mix (slow end-over-end) for at least 15 minutes.

c. Remove the liquid.

3. Washing

a. Add 500 μl Binding Buffer containing 0.5 M NaCl; Remove the liquid.

b. Add 500 μl Binding Buffer; Remove the liquid.

4. Target Binding

a. Add sample diluted in binding buffer.

b. Incubate with slow end-over-end mixing for up to 60 minutes.

c. Remove and collect unbound fraction.

5. Washing ( perform three times)

a. Add 500 μl wash buffer

b. Remove liquid (save washes to troubleshoot)

6. Elution (perform three times)

a. Add 2 volumes elution buffer (vs. bead volume).

b. Completely resuspend beads and incubate at least 2 minutes.

c. Remove and collect elution fraction.

RECOMMENDED BUFFER EXAMPLES:

Binding buffer: 50 mM Tris, 150 mM NaCl, pH 7.5

Wash buffer: 50 mM Tris, 150 mM NaCl, pH 7.5 (or add 1% Octylglucoside to this buffer) (Could also try 1X PBS as both binding and wash buffer)

Elution buffer: 0.1 M -0.2 M Glycine pH 2.5-3.1 (or 0.1 M citric acid, pH 2.5-3.1 or 2.5 % Acetic Acid)