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| Size/SKU | Availability | Price |
|---|---|---|
2 μg | Available to ship TODAYfromMILWAUKEE | $230.00 |
3 x 2 μg | Available to ship TODAYfromMILWAUKEE | $624.00 |
$230.00
Available to ship TODAYDetails
assay
≥90% (SDS-PAGE)
form
lyophilized
specific activity
≥20000 units/mg protein
mol wt
27 kDa
packaging
pkg of 2 μg (11420488001), pkg of 3 × 2 μg (11054589001)
manufacturer/tradename
Roche
optimum pH
7.0-8.0
pH range
6.5-9.0
storage temp.
2-8°C
General description
The wild type of Pseudomonas fragi segregates a metalloprotease, which cleaves peptide bonds N-terminally at small hydrophilic amino acids. A mutant of this strain produces the Endoproteinase Asp-N Sequencing Grade, which is isolated as a highly purified and specific protease.
Application
Biochem/physiol Actions
Endoproteinase Asp-N Sequencing Grade is a metalloprotease. In phosphate, acetate, or Tris buffers at pH 6.0–8.5, it specifically cleaves peptide bonds N-terminally at aspartic and cysteic acid. If cysteine is reduced or alkylated, only -↓-Asp-X is cleaved.
The specificity of Endoproteinase Asp-N Sequencing Grade is verified with glucagon as a substrate. High concentrations of Endoproteinase Asp-N Sequencing Grade (1 part by weight enzyme with 10 parts by weight glucagon) are incubated to detect traces of impurities like contaminating proteases.
Heat inactivation: The protease is stopped by boiling for 5 minutes.
The specificity of Endoproteinase Asp-N Sequencing Grade is verified with glucagon as a substrate. High concentrations of Endoproteinase Asp-N Sequencing Grade (1 part by weight enzyme with 10 parts by weight glucagon) are incubated to detect traces of impurities like contaminating proteases.
Heat inactivation: The protease is stopped by boiling for 5 minutes.
Preparation Note
Working concentration: 2 μg /50 μl double-dist. water
One vial (= 2 μg enzyme) is dissolved in 50 μl double-dist. water resulting in a buffer concentration of 10 mM Tris-HCl pH 7.5.
Working solution: Recommended solvent is double-distilled water.
Storage conditions (working solution): -15 to -25 °C
A solution in double-dist. water is stable for 1 week at maximum at 2 to 8 °C and for about 1 month at -15 to -25 °C; repeated thawing should be avoided.
One vial (= 2 μg enzyme) is dissolved in 50 μl double-dist. water resulting in a buffer concentration of 10 mM Tris-HCl pH 7.5.
Working solution: Recommended solvent is double-distilled water.
Storage conditions (working solution): -15 to -25 °C
A solution in double-dist. water is stable for 1 week at maximum at 2 to 8 °C and for about 1 month at -15 to -25 °C; repeated thawing should be avoided.
Lyophilized Endoproteinase Asp-N sequencing grade is reconstituted in 50 μl double-dist. water. This results in a buffer concentration of 10 mM Tris-HCl, pH 7.5. In order to avoid autolysis the incubation temperature should not exceed 37 °C.
Other Notes
For life science research only. Not for use in diagnostic procedures.
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This Item | |||
|---|---|---|---|
| assay ≥90% (SDS-PAGE) | assay - | assay - | assay ≥90% (SDS-PAGE) |
| specific activity ≥20000 units/mg protein | specific activity ≥15 units/mg protein | specific activity ≥200 units/mg protein (at 25 °C, with Chromozym PL) | specific activity ≥20,000 units/mg protein |
| form lyophilized | form lyophilized (salt-free) | form lyophilized | form lyophilized solid |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C |
| mol wt 27 kDa | mol wt Mr 27 kDa | mol wt 33 kDa by SDS-PAGE (reducing) | mol wt - |
| optimum pH 7.0-8.0 | optimum pH 4.0-7.8 | optimum pH 8.5-8.8 | optimum pH - |
Storage Class
11 - Combustible Solids
flash_point_f
does not flash
flash_point_c
does not flash
wgk
WGK 3
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