Collagen fixation and staining kit

Extracellular matrix including collagen fibers is particularly difficult to fix and label with antibodies, largely because the strong molecular interactions within the matrix block the antibody access. BiCell Scientific has found that a combination of dehydrant and hydrogen bond breaker can effectively disrupt the molecular interactions in the extracellular matrix and expose the individual protein constituent for antibody recognition.

Dehydrants such as methanol and acetone remove and replace free water in the cell and tissue, and cause a change in the molecular interaction landscape of the extracellular matrix by destabilizing the hydrophobic interaction. Hydrophobic areas, frequently found on the inside of protein molecules, are exposed due to the repulsion of water, allowing antibody access to otherwise inaccessible protein domains.

Hydrogen bond breakers such as urea and acid can disrupt the hydrogen bonds made both between and within protein molecules that are vital for the high-order oligomeric structure of the extracellular matrix, such as collagen fiber. As a result, protein oligomers are dissociated into monomers, which greatly facilitates the antibody access and binding.