Alkaline phosphatase (AP) is an enzyme that catalyzes the hydrolysis of phosphate groups, enabling sensitive colorimetric or chemiluminescent detection when conjugated to an antibody. For anti-rat reagents, AP is typically covalently linked to the immunoglobulin via protein cross-linking with 0.2% glutaraldehyde. The affinity-isolated antibody is obtained from anti-rat IgG antiserum by immunospecific purification prior to conjugation, ensuring that the enzyme is attached to a specific immunoglobulin fraction. The resulting conjugate is supplied in a buffered solution often containing bovine serum albumin (BSA) as a stabilizer and magnesium chloride to maintain enzymatic activity.

Applications

AP-conjugated anti-rat reagents are validated across several key immunoassay platforms. They are used in Western blotting for detecting rat proteins on membranes, ELISA for quantifying rat immunoglobulins or specific antigens, and immunohistochemistry (IHC) for visualizing target proteins in rat tissue sections. For immunohistochemistry, these reagents enable spatial localization of antigens in both frozen and paraffin-embedded tissues. Working dilutions vary by application and product, with representative ranges of approximately 1:5,000–1:50,000 for ELISA and Western blotting, and 1:200–1:1,000 for IHC, depending on the specific conjugate and detection system.