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CIAP (Calf Intestinal Alkaline Phosphatase), 20 U/μL
Calf Intestinal Alkaline Phosphatase (CIAP) is a phosphomonoesterase that removes 3´ and 5´ phosphates from DNA and RNA.Concentration: 20 units/μLApplications:Read more
| Catalog Number | Quantity |
|---|---|
| 18009019 | 1000 U |
Catalog number 18009019
Price (USD)
149.65
Online Exclusive
158.00Save 8.35 (5%)
Each
-
Quantity:
1000 U
Price (USD)
149.65
Online Exclusive
158.00Save 8.35 (5%)
Each
Calf Intestinal Alkaline Phosphatase (CIAP) is a phosphomonoesterase that removes 3´ and 5´ phosphates from DNA and RNA.
Concentration: 20 units/μL
Applications: Dephosphorylation of 5´-phosphorylated termini of vector DNA to prevent self-ligation (1). Dephosphorylation of 5´ termini of nucleic acids prior to forward reaction with kinase.
Source: Purified from calf intestinal mucosa.
Performance and Quality Testing: Endodeoxyribonuclease, 3´ exodeoxyribonuclease, and ribonuclease assays; dephosphorylation efficiency measured in a transformation assay.
Unit Definition: One unit hydrolyzes 1 μmol of 4-nitrophenyl phosphate in 1 min. at 37°C.
Unit Reaction Conditions: 1 M diethanolamine buffer, 10 mM 4-nitrophenyl phosphate, 0.25 mM MgCl2 (pH 9.8) in 900 μl for 10 min. at 37°C.
Concentration: 20 units/μL
Applications: Dephosphorylation of 5´-phosphorylated termini of vector DNA to prevent self-ligation (1). Dephosphorylation of 5´ termini of nucleic acids prior to forward reaction with kinase.
Source: Purified from calf intestinal mucosa.
Performance and Quality Testing: Endodeoxyribonuclease, 3´ exodeoxyribonuclease, and ribonuclease assays; dephosphorylation efficiency measured in a transformation assay.
Unit Definition: One unit hydrolyzes 1 μmol of 4-nitrophenyl phosphate in 1 min. at 37°C.
Unit Reaction Conditions: 1 M diethanolamine buffer, 10 mM 4-nitrophenyl phosphate, 0.25 mM MgCl2 (pH 9.8) in 900 μl for 10 min. at 37°C.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeAlkaline Phosphatase
Quantity1000 U
Shipping ConditionApproved for shipment on Wet or Dry Ice
Concentration20 U/μL
EnzymeCIP
Unit SizeEach
Contents & Storage
Calf Intestinal Alkaline Phosphatase (20 units/μL) is supplied with a vial of 10X dephosphorylation buffer [500 mM Tris-HCl (pH 8.5), 1 mM EDTA], vial of dilution buffer [50% glycerol, 25 mM Tris-HCl (pH 7.6), 1 mM MgCl2 , and 0.1 mM ZnCl2 ]. Store Calf Intestinal Alkaline Phosphatase at -20°C.
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Lot #Certificate TypeDateCatalog Number(s)
3188376Certificate of AnalysisApr 30, 202518009019
3209037Certificate of AnalysisApr 23, 202518009019
3177396Certificate of AnalysisMar 31, 202518009019
3136781Certificate of AnalysisFeb 28, 202518009019
3102174Certificate of AnalysisJan 22, 202518009019
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Frequently asked questions (FAQs)
Dephosphorylating the vector to decrease background can be achieved with:
Heat Inactivated Alkaline Phosphatase, or Calf Intestinal Alkalline Phosphatase (available as 1 unit/µL or 20 units/µL)
If you need to create blunt phosphorylated DNA ends (“polishing” the ends), you can use:
DNA End Repair Mix or T4 DNA polymerase or Klenow Fragment (large fragment) of E. coli DNA polymerase to generate blunt ends due to their 5' to 3' DNA polymerase activity (filling-in of 5' overhangs) and 3' to 5' exonuclease activity (chewing back of 3' overhangs).
CIP/CIAP or BAP can dephosphorylate the 5' ends of DNA/RNA. T4 Polynucleotide Kinase can add back phosphate groups.
Alkaline phosphatases are used to dephosphorylate the 5' ends of DNA. In cloning, it is used to prevent self-ligation of vector DNA. Standard ligation of DNA with ligase requires a 5' phosphate to be present on at least one of the ends being joined. When a DNA insert containing phosphates on both 5' termini is added to a dephosphorylated vector, the insert will be efficiently ligated into the vector, but the vector will not be able to self-ligate. Thus, dephosphorylation of vector lowers the number of background colonies containing vector without insert.