2.1. Single-Point Attachment: Thermal Unfolding

2.1.1. Activation of Silica Beads

1. Controlled pore carrier (CPC)-silica beads, aminopropyl derivative, pore size 500 Ä, 30-45 mesh (Fluka,, 10 g (dry weight; see Note 1).

2. Washing buffer: phosphate buffer 100 mM, pH 7.0.

3. 10% (v/v) Aqueous glutaraldehyde (di-aldehyde) solution.

2.1.2. Protein Immobilization

1. Protein: ribonuclease A (RNase, Sigma, Type XII A, molecular weight [MW] 13.700).

2. Immobilization buffer: 100 mMphosphate buffer, pH 7.0.

3. Glutaraldehyde activated CPC silica beads.

2.1.3. Number of Linkages Between Protein and Support

1. Protein: ribonuclease A RNase (Sigma, Type XII A).

2. Buffer: 100 mM sodium phosphate, pH 7.0.

3. Aqueous valeraldehyde (monoaldehyde) solution: 10% (v/v).

4. Solution of trinitrobenzenesulfonic acid, according to standard literature conditions (15).

Was this article helpful?

0 0
Detoxify the Body

Detoxify the Body

Need to Detoxify? Discover The Secrets to Detox Your Body The Quick & Easy Way at Home! Too much partying got you feeling bad about yourself? Or perhaps you want to lose weight and have tried everything under the sun?

Get My Free Ebook

Post a comment