Heat Inactivated Alkaline Phosphatase is a phosphomonoesterase that hydrolyzes 3’ and 5’ phosphates from DNA and RNA. It is suitable for removing 5’ phosphates prior to end-labeling and for dephosphorylating vectors prior to insert ligation.  It can also be used to dephosphorylate proteins.

Features of Heat Inactivated Alkaline Phosphatase:

  • Removes 5’ phosphate groups from DNA, RNA, and protein
  • Dephosphorylates in 15 minutes at 37 °C
  • Heat inactivated in 5 minutes at 65 °C

Reduce Vector Background, Clean Up PCR Reactions, Dephosphorylate Proteins

Since the 5’ phosphate group is required for DNA ligation, treatment with Heat Inactivated Alkaline Phosphatase prevents vector self-ligation and recircularization, resulting in decreased vector background when cloning.  In addition, Heat Inactivated Alkaline Phosphatase can be used to remove pyrophosphate from PCR reactions and to dephosphorylate proteins, though specific reaction conditions will vary depending on the protein. It is not recommended for use in removing dNTPs.

Easily Inactivated So Removal Is Not Necessary

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Heat Inactivated Alkaline Phosphatase is completely inactivated in 20 minutes at 65°C, so no phenol/chloroform extraction or column purification is required to remove the enzyme prior to ligation. This means DNA fragments added to the reaction mix for ligation are not treated with residual phosphatase and will be ligated to the dephosphorylated vector DNA.

For research use only. Not intended for human or animal therapeutic or diagnostic use.