Takara Edman degradation and protein sequencing: Pfu N-acetyl Deblocking Aminopeptidase (Ac-DAP)

More Information

Used for the liberation of N-terminal formyl, acetyl, and myristyl blocking groups from proteins

Components

• Buffer (5X)
  Once thawed, the buffer should be stored at 4°C. Avoid additional freeze-thaw cycles.

|     |     |
| --- | --- |
| Volume: | 1 ml |
| Components: | 250 mM _N_\-Ethylmorpholine-AcOH buffer (pH 8.0, 0.5 mM CoCl2) |
| Once thawed, the buffer should be stored at 4°C. Avoid additional freeze-thaw cycles. |     |

Source

Recombinant yeast containing the gene encoding Pyrococcus furiosus Ac-DAP.

Storage

−20°C. Once thawed, the enzyme solution should be stored at 4°C. The thawed enzyme and buffer are stable for at least 3 months at 4°C.

Notes

This enzyme cannot react with proteins containing higher order structures; protein samples should be denatured by chemical procedures such as carboxymethylation. This enzyme cannot react with proteins blotted onto PVDF membranes.

Properties

• Specific activity: >8 units/mg
• Protein concentration: 1 mg/ml
• Molecular weight: 38,639 Da (mass spectrometry), ~451 kDa (sedimentation equilibrium method)
• Activator: CoCl2
• Inhibitor: Amastatin, EDTA
• Optimum pH: 6.5–9.0
• Optimum Temperature: 85–95°C
• Thermal stability: Enzyme retains 100% activity after 48 hr at 50°C in the supplied buffer (pH 8.0, 0.1 mM Co2+)
• N-terminal amino acid sequence: Ac-MDDYELLKKVVEADGV...

Form

In a solution of 50 mM N-Ethylmorpholine-AcOH buffer (pH 8.0)

Definition of activity

One unit of activity corresponds to the amount of enzyme required to hydrolyze 1 µmol of leucine-p-nitroanilide at 75°C and pH 8.0 in one minute.

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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