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Cat. No.: EPAP-100 (for 100U)

Cat. No.: EPAP-500 (for 500U)

Cat. No.: EPAP-2500 (for 2500U)

Cat. No.: EPAP-10k (for 10KU)

Description

E. coli Poly(A) Polymerase, also known as E. coli PAP, PAP enzyme, Poly(A) polymerase, or Poly(A) tailing enzyme, facilitates the addition of AMP, derived from ATP, to the 3' end of single-stranded RNA in a template-independent manner, leading to the formation of a Poly(A) tail. E. coli Poly(A) Polymerase can employ various single-stranded RNAs as substrate RNAs. However, double-stranded RNA and short oligonucleotides are not recommended for this reaction, and DNA cannot be utilized as a substrate. Experimental evidence shows that RNA synthesized with 15 nucleotides can effectively serve as a substrate for E. coli Poly(A) Polymerase, enabling the catalysis of Poly(A) tail formation in the presence of ATP. The Poly(A) tailing reaction catalyzed by E. coli Poly(A) Polymerase exclusively employs ATP and cannot utilize ADP or dATP. Moreover, when using CTP and UTP, their incorporation is less than 5% compared to ATP, and GTP cannot be added to the 3' end of RNA at all.

Application

• Labeling RNA with radiolabeled ATP or cordycepin.
• Adding a Poly(A) tail to RNA for reverse transcription cloning or affinity purification.
• Increasing mRNA stability to enhance translation efficiency of RNA transfected into eukaryotic cells.

Source

Recombinant E. coli strain carrying the Poly(A) polymerase gene cloned from Escherichia coli.

Activity Definition

One unit is defined as the amount of enzyme that incorporates 1 nmol of AMP into RNA in a 20 µl volume within 10 minutes at 37°C.

Purity

Free from DNase, RNase, and phosphatases.

Enzyme Storage Buffer

20 mM Tris-HCl (pH 7.5, 25°C), 500 mM NaCl, 1 mM DTT, 1 mM EDTA, 50% (v/v) Glycerol, 0.1% (w/v) Triton X-100.

10X Reaction Buffer

500 mM Tris-HCl (pH 8.1, 25°C), 2.5 M NaCl, 100 mM MgCl2.

Inactivation or Inhibition

Add EDTA to a final concentration of 10 mM or heat at 65°C for 20 minutes.

Storage

The minimum shelf life is 2 years at -20°C.

Note

• The 10X PAP Reaction Buffer does not contain ATP.
• The dosage of E. coli Poly(A) Polymerase primarily affects the length of the desired product. The enzyme dosage can be adjusted accordingly based on the desired product length. If longer products are desired, the enzyme dosage can be increased or the reaction time can be extended.
• This product is intended for scientific research purposes by professionals only. It is not to be used for clinical diagnosis or treatment, nor for food or drug purposes. It should not be stored in residential areas.
• For your safety and health, please wear lab attire and disposable gloves while handling.

Only for research and not intended for treatment of humans or animals

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