What Reagents Are Required for In Vitro Cross-Linking of Two Purified Proteins
Chemical cross-linkers are commonly employed for in vitro protein cross-linking. The choice of reagent depends on the characteristics of the proteins involved and the specific goals of the experiment. Commonly used protein cross-linkers include:
1. BS3 (Bis(sulfosuccinimidyl)suberate)
BS3 is a water-soluble cross-linker that targets lysine residues for cross-linking.
2. DSS (Disuccinimidyl suberate)
DSS is structurally similar to BS3 but is water-insoluble, making it suitable for non-aqueous environments.
3. EDC (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide)
EDC is a zero-length cross-linker that facilitates the formation of covalent bonds between carboxyl and amino groups.
4. Glutaraldehyde
Glutaraldehyde is capable of forming extensive cross-linking networks through reactions with amino groups.
5. Sulfo-EGS (Ethylene glycol bis(sulfosuccinimidyl succinate))
Sulfo-EGS is highly water-soluble and features a longer spacer arm, enabling cross-linking over greater molecular distances.
When selecting a suitable cross-linker, one must take into account the physicochemical properties of the reagent—such as spacer length, solubility, and reactivity—as well as the properties of the target proteins and relevant experimental conditions, including pH and temperature.
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