The Protein Man's Blog | A Discussion of Protein Research

The Top 8 Applications for Protein Cross-Linkers and Why You Need Them (Part 1)

Posted by The Protein Man on Nov 26, 2013 5:00:00 AM
The Protein Man

Question:

What are the applications of protein cross linkers?  

The Protein Man Says:

Protein cross-linking generally refers to the process of chemically joining two or more molecules or polymer chains in an effort to check and investigate protein-protein interactions. Cross-linking agents or "cross-linkers" as they are fondly called by most researchers, have the ability to covalently bind with two or more molecules or functional groups since they contain at least two reactive ends that can attach themselves to several functional groups such as primary amines, carboxyls, sulfhydryls, carbohydrates and carboxylic acids.

protein cross linking

Cross-Linking Applications

Due to their unique nature, cross-linking agents can be used in the following applications:

  • Structural and functional studies

  • Protein and receptor interactions

  • Preparation of immunogens and immunotoxins

  • Cell membrane and cell surface studies

  • Solid-phase immobilization

  • Protein-protein conjugation

  • Protein to DNA/RNA cross-linking

  • Reactive group transfer

Structural and functional studies

Cross-linking agents can be used to investigate the structure, subunit composition and various folding patterns of a particular protein and stabilize the conformational changes that proteins undergo. You can also experiment with different lengths of identical cross-linkers to quantify the molecular distance between functional groups within the protein structure. Just keep in mind that cross-linkers with short to medium spacer arms are suitable for intramolecular cross-linking while those with longer spacer arms are best for intermolecular cross-linking.

Protein and receptor interactions

Protein cross-linkers can also help you establish the association between proteins and the interactions that exist between ligands and receptors. You can use a panel of similar cross-linkers with varying spacer arm lengths to achieve this purpose, especially if you know the distance between two potential molecules. Heterobifunctional cross-linkers with one non-specific and photo-activatable end can be particularly useful for protein interaction analysis.

Preparation of immunogens and immunotoxins

Cross-linking agents can also be used in the production of antibodies and immunotoxins. Sulfhydryl-amine reactive heterobifunctional cross-linkers and carbodiimides are ideally used in the preparation of antibodies while cleavable disulfide-containing cross-linkers are found to be more suitable in the preparation of immunotoxins.

We will take a look at some of the other important applications of protein cross-linking in our next post so please watch out for it!

Topics: Cross-Linkers

Want more Protein Man blogs?

Purification_resins
Ellyn Daugherty's Biotechnology: Science for the New Millennium

CB™ PROTEIN ASSAY: A Bradford Protein Assay

CB Protein Assay Graph

An improved Coomassie Dye based protein assay based on the Bradford Protein Assay. This assay is suitable for the simple and rapid estimation of protein concentration. This assay is based on a single Coomassie dye based reagent. The binding of protein to the dye results in a change of color from brown to blue. The change in color density is proportional to protein concentration. Protein estimation can be performed using as little as 0.5µg protein.

Features

  • Sensitivity: Linear responses over the range of 0.5µg-50µg protein
  • Flexible Protocols: Suitable for tube or Titer plate assays
  • Ready to use assay reagents and no preparation required
  • Long shelf life, stable for 12 months
Click for CB Protein Assay