Description
Calf Intestinal Alkaline Phosphatase (CIAP) – 20 U/µL | ZELLX®
Product Overview
ZELLX® Calf Intestinal Alkaline Phosphatase (CIAP) is a premium-grade molecular biology enzyme derived from calf intestinal mucosa, one of the tissue-specific mammalian alkaline phosphatase isoenzymes. Alkaline phosphatases belong to a conserved superfamily of metallohydrolases that catalyze the hydrolytic removal of phosphate groups from a broad range of substrates, including nucleic acids.
ZELLX® CIAP is highly purified to remove contaminating nucleases and proteases, ensuring reliable performance and minimal background activity in sensitive downstream applications. The enzyme efficiently catalyzes the removal of 5′-phosphate groups from DNA and RNA, including ribo- and deoxyribonucleoside triphosphates, providing precise control over ligation reactions and improving cloning efficiency.
Supplied at a high activity concentration of 20 U/µL, ZELLX® CIAP enables rapid and efficient dephosphorylation using minimal enzyme volumes, reducing reagent consumption and cost per experiment. The enzyme acts effectively on 5′ protruding ends, 5′ recessed ends, and blunt-ended DNA, making it a versatile tool for a wide range of molecular biology workflows.
Manufactured under strict quality standards, ZELLX® CIAP delivers excellent reproducibility and batch-to-batch consistency, making it suitable for routine and advanced applications in research, diagnostics, and biotechnology laboratories. Designed for compatibility with standard laboratory buffers and workflows, it integrates seamlessly into existing protocols without the need for complex optimization.
ZELLX® CIAP is widely used for vector dephosphorylation, cloning, end-labeling, and general nucleic-acid modification workflows, where prevention of plasmid self-ligation and reliable downstream performance are critical.
Unit Definition
One unit (U) of Calf Intestinal Alkaline Phosphatase is defined as the amount of enzyme required to hydrolyze 1 µmol of p-nitrophenyl phosphate (pNPP) per minute in a 1 mL reaction volume at 37 °C, under specified assay conditions.
Product Specifications:
Catalog Number: ZXB-06-202
DNA Dephosphorylation Protocol Using ZELLX® Calf Intestinal Alkaline Phosphatase (CIAP)
Materials Required
- DNA sample
- ZELLX® Calf Intestinal Alkaline Phosphatase (CIAP), 20 U/µL
- 1× Alkaline Phosphatase Reaction Buffer
- Nuclease-free water
Procedure
- Prepare the DNA substrate
Dissolve DNA in 1× Alkaline Phosphatase Reaction Buffer at a concentration of 0.5 µg DNA per 10 µL reaction volume.
- Add CIAP enzyme
Add ZELLX® CIAP according to the DNA end structure:
- 5′ overhang DNA: 0.1 U CIAP per pmol of DNA ends
- 3′ overhang or blunt-end DNA: 1 U CIAP per pmol of DNA ends
- Incubation
Mix gently and incubate at 37 °C for 60 minutes.
- DNA purification
Purify the DNA using agarose gel purification, spin-column purification, or phenol–chloroform extraction.
Heat Inactivation (Optional)
Greater than 95% of CIAP activity can be inactivated by heating the reaction to 75 °C for 10 minutes in the presence of 5 mM EDTA.
If DNA is purified by phenol extraction or gel purification, heat inactivation is not required.
