What does SCAM mean in LABORATORY

SCAM provides researchers with rapid and reliable data, as it typically requires few steps with minimal sample preparation before experiments can be conducted. In addition, it does not require changes to the protein's original structure and can be used in combination with other techniques to provide more comprehensive analysis.

The basis of SCAM is that a reactive group attached to cysteine residues can be modified by adding a probe molecule, which serves as a tag or label. By being able to observe these tags, researchers can gain insight into which parts of the protein structure are exposed on its surface.

For SCAM experiments, fluorescent molecules or enzymes are typically used as probes due to their stable covalent attachment when bound to cysteines through functional groups like maleimides or thiols.

Purified protein samples need to be cleaved with trypsin prior to labeling with probes in order for them to have accessible cysteines available for binding. Crude cell lysates also may be used in some cases without further preparation if they contain sufficient amounts of active proteases participating in enzymatic reactions known as autocleavage.

While there are various approaches available for determining the accessibility of surfaces presented by proteins, such as chemical or hydroxyl radical probing or hydrogen/deuterium exchange (HDX), none offer the sensitivity and specificity provided by SCAM.

When conducting experiments, users should keep in mind that reactions between cysteines and probes may vary depending on factors such as pH range and temperature conditions of the reaction mixture since these affect how well probes bind when added during labeling protocols. Additionally, large molecular size of certain probes may present certain obstacles depending on the target protein being studied.

When preparing samples for precisely measuring surface accessibilities via this method it is important that any existing disulfide linkages remain intact because breaking them may change both positioning and orientation of residues created affecting data accuracy collected from analyses performed thereafter.