Crosslinking Reagents

Crosslinking, also known as bioconjugation, is achieved with the use of bifunctional reagents containing reactive end groups that respond to functional groups. There are four functional groups that have been found to be successful cross-linking targets. These functional groups are primary amines, carboxyls, sulfhydryls and carbonyls. The bifunctional reagents are characterized as either homobifunctional or heterobifunctional. Together these allow for the formation of intermolecular and intramolecular crosslinking enabling researchers to study the interactions and proximity of neighboring proteins.

Heterobifunctional Crosslinking Reagents

Heterobifunctional reagents consist of 2 different reactive groups on either end of a spacer arm. These reagents can provide more specific information about the protein interactions and their proximity to one another. This is a result of their ability to perform multiple step cross-linking reactions. In the first step, a protein is modified with one reactive group of the heterobifunctional reagent; the remaining free reagent is removed. In the second step, the modified protein is mixed with a second protein, which is then allowed to react with modifier group at the other end of the reagent. The advantage of having the 2 steps in the conjugation process is that unwanted polymerization or self-linking can be minimized during the free agent removal step.

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B8271 Bromoacetic acid N-hydroxysuccinimide ester ≥95%, powder
M2786 3-Maleimidobenzoic acid N-hydroxysuccinimide ester crystalline
M5525 4-(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester ≥98%, powder
P3415 3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester ≥95%, powder

Homobifunctional Crosslinking Reagents

Homobifunctional reagents consist of identical reactive groups on either end of a spacer arm. They are used for single step conjugation and are extremely useful when evaluating protein interactions.
There are different types of homobifunctional reagents. Homobifunctional imidoesters are useful in determining the distances between linked residues on surfaces protein that are close to each other and reversible homobifunctional crosslinkers, such as N-hydroxysuccinimide (NHS) esters, are used for identification of unknown interacting proteins.

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D8388 Dimethyl pimelimidate dihydrochloride powder
D3669 3,3′-Dithiodipropionic acid di(N-hydroxysuccinimide ester) powder
S5799 Suberic acid bis(3-sulfo-N-hydroxysuccinimide ester) sodium salt ≥95% (H-NMR), powder