Alkaline Phosphatase Buffer-2 (pH 9.5, 1×) is widely used in molecular biology and biochemistry for enzymatic reactions involving alkaline phosphatase. It provides an optimal pH environment for the enzyme to function efficiently, making it essential for applications such as dephosphorylation of DNA, RNA, and proteins. This buffer is commonly used in cloning experiments to prevent self-ligation of plasmid vectors by removing 5' phosphate groups. It is also utilized in immunoassays, such as ELISA, where alkaline phosphatase is employed as a reporter enzyme. Additionally, the buffer is suitable for histochemical staining to detect alkaline phosphatase activity in tissue samples. Its stable pH ensures consistent performance in various diagnostic and research applications.