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Cat. No.: BAP-50 (for 50U)

Cat. No.: BAP-250 (for 250U)

Description

Bacterial Alkaline Phosphatase (BAP) is a novel alkaline phosphatase that can non-specifically catalyze the dephosphorylation of almost all phosphomonoesters and degrade the pyrophosphate bond of ATP. It can achieve 100% activity in various endonucleases and PCR buffers within 30 minutes, completing the dephosphorylation of various types of DNA and RNA 5' ends. It maintains high activity at temperatures ranging from 37-65ºC.

Alkaline Phosphatase (AP/ALP/AKP/ALKP/ALPase/Alk Phos), often referred to as alkaline phosphatase (EC 3.1.3.1), is a type of hydrolase. It removes phosphate groups from substrate molecules by hydrolyzing phosphomonoesters, producing phosphate ions and free hydroxyl groups. Its dephosphorylation substrates include nucleotides, proteins, and alkaloids, and it is most effective under alkaline conditions. This enzyme is a group of isoenzymes. Commonly, calf intestinal alkaline phosphatase (CIAP/CIP) is widely used for labeling secondary antibodies for protein and nucleic acid detection, as well as for the dephosphorylation of DNA or RNA 5' and 3' ends, particularly to prevent plasmid self-ligation during cloning.

Plasmids with 5' phosphate groups generated by restriction enzyme digestion can use Bacterial Alkaline Phosphatase to remove the 5' phosphate groups to prevent self-ligation during gene cloning. Dephosphorylated 5' ends of plasmids cannot self-ligate.

Features

• Broad substrate specificity: Non-specifically catalyzes the dephosphorylation of almost all phosphomonoesters.
• High thermal stability: Retains activity for one hour at 65ºC and is only temporarily inactivated by heating at 100ºC with chelating agents, requiring phenol treatment for complete inactivation.
• Compatibility: Achieves 100% activity in various endonucleases and almost all PCR buffers, and can be used simultaneously with plasmid DNA endonuclease digestion.
• Simple operation: Uniform procedure for dephosphorylation of 5' or 3' overhangs and blunt ends of various DNA.

Applications

• Prevents self-ligation of vector or DNA fragments by removing 5' phosphate groups.
• Simultaneous endonuclease digestion and 5' dephosphorylation of plasmid DNA.
• Prepares templates for 5' end phosphate labeling.
• Removes 5' or 3' phosphate groups from DNA or RNA.
• Dephosphorylates phosphoserine, phosphothreonine, and phosphotyrosine residues in proteins.

Source

Purified from E. coli recombinant strains carrying the phoA gene.

Enzyme Activity Definition

One unit is the amount of enzyme that produces 1 μmol of p-nitrophenol per minute at 25ºC and pH 8.0 with p-nitrophenyl phosphate as the substrate. If the enzyme activity of this product is defined as "One unit hydrolyzes 1 nmol of ATP in 30 minutes at 37ºC," one unit of enzyme activity is equivalent to approximately 1000 units, with product packaging equivalent to approximately 50 kU and 200 kU.

Purity

Free of DNA exonucleases and endonucleases, and RNAse.

Inactivation or Inhibition

Bacterial Alkaline Phosphatase can be temporarily inactivated by heating at 100ºC with chelating agents and can regain activity when the temperature drops to room temperature. Phenol can completely inactivate it. Inorganic phosphate significantly inhibits Bacterial Alkaline Phosphatase.

Storage

Store at -20ºC.

Precautions

• The enzyme should be kept in an ice box or ice bath while in use, and should be placed back at -20ºC immediately after use.
• This product is for scientific research purposes only, by professional personnel. It is not for clinical diagnosis or treatment, not for use in food or drugs, and should not be stored in common residential areas.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.

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