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Recombinant Human Cathepsin E Protein, CF

Catalog #: 1294-AS
3 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
1294-AS-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (3)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Cathepsin E Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to cleave the fluorogenic peptide substrate, Mca-PLGL-Dpa-AR-NH2 (Catalog # ES001). The specific activity is >1,500 pmol/min/µg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Cathepsin E protein
Gln18-Pro396, with a C-terminal 10-His tag
Accession #
NP_001901
N-terminal Sequence
Analysis
Ile54 & Gln18 (predicted)
Structure / Form
Pro and mature forms
Predicted Molecular Mass
42 kDa
SDS-PAGE
48 kDa, reducing conditions

Product Datasheets

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1294-AS

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

1294-AS

Formulation Lyophilized from a 0.2 μm filtered solution in MES and NaCl.
Reconstitution Reconstitute at 100 μg/mL in sterile 25 mM MES, 150 mM NaCl, pH 6.5.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Assay Procedure

Materials
  • Assay Buffer: 0.1 M NaOAc, 0.5 M NaCl, pH 3.5
  • Recombinant Human Cathepsin E (rhCathepsin E) (Catalog # 1294-AS)
  • Fluorogenic Peptide Substrate I: MCA-Pro-Leu-Gly-Leu-DPA-Ala-Arg-NH2 (Catalog # ES001)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin E to 1.0 µg/mL in Assay Buffer.
  2. Incubate at room temperature for 30 minutes (required to fully activate).
  3. Dilute activated rhCathepsin E to 0.2 ng/µL in Assay Buffer.
  4. Dilute Substrate to 40 µM in Assay Buffer.
  5. Load 50 µL of the 0.2 ng/µL rhCathepsin E into a black well plate, and start the reaction by adding 50 µL of 40 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 40 µM Substrate without any rhCathepsin E.
  6. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard MCA-Pro-Leu-OH (Bachem, Catalog # M-1975).

Per Well:
  • rhCathepsin E: 0.01 µg
  • Substrate: 20 µM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Cathepsin E

Cathepsin E is an intracellular aspartic protease of the pepsin family (1). Unlike Cathepsin D, another member of the same family and a lysosomal protease with relatively ubiquitous distribution, Cathepsin E is not a lysosomal enzyme and has a limited cell and tissue distribution. However, both Cathepsin D and E play an important role in the degradation of proteins, the generation of bioactive proteins, and antigen processing (2). Both enzymes are efficient in cleaving Swedish mutant of amyloid precursor protein (APP) at the beta  site but show almost no reactivity with wild-type APP (3). Human Cathepsin E is synthesized as a precursor protein, consisting of a signal peptide (residues 1‑17), a propeptide (residues 18‑53), and a mature chain (residues 54‑396) (4).

References
  1. Kay, J. and P.J. Tatnell (2004) in Handbook of Proteolytic Enzymes (Barrett, A.J. et al. eds.), p. 33, Academic Press, San Diego.
  2. Tsukuba, T. et al. (2000) Mol. Cells 10:601.
  3. Gruninger-Leitch, F. et al. (2000) Nat. Biotechnol. 18:66.
  4. Azuma, T. et al. (1989) J. Biol. Chem. 264:16748.
Entrez Gene IDs
1510 (Human); 13034 (Mouse)
Alternate Names
CATE; Cathepsin E; CTSE; EC 3.4.23; EC 3.4.23.34; erythrocyte membrane aspartic proteinase; slow-moving proteinase

Citations for Recombinant Human Cathepsin E Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Absence of the CXCR4 antagonist EPI-X4 from pharmaceutical human serum albumin preparations
    Authors: A Gilg, M Harms, LR Olari, AK Urbanowitz, H Bonig, J Münch
    Journal of Translational Medicine, 2021-05-03;19(1):190.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. Global protease activity profiling provides differential diagnosis of pancreatic cysts
    Authors: SL Ivry, JM Sharib, DA Dominguez, N Roy, SE Hatcher, M Yip-Schnei, CM Schmidt, RE Brand, WG Park, M Hebrok, G Kim, AJ O'Donoghue, KS Kirkwood, CS Craik
    Clin. Cancer Res., 2017-04-19;0(0):.
    Applications: Bioassay
  3. Cathepsin D-mediated proteolysis of apolipoprotein E: possible role in Alzheimer's disease.
    Authors: Zhou W, Scott SA, Shelton SB, Crutcher KA
    Neuroscience, 2006-09-25;143(3):689-701.
    Species: Human
    Sample Types: Tissue Homogenates
    Applications: Bioassay

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