Recombinant Human Cathepsin X/Z/P Protein, CF

R&D Systems | Catalog # 934-CY

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Cathepsin X/Z/P Protein (934-CY)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

NS0

Accession Number

Q9UBR2

Structure / Form

Pro form

Applications

Enzyme Activity
View all Cathepsin X/Z/P Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Cathepsin X/Z/P protein
Ala21-Val303 (Gly23Val & Ser48Thr), with a C-terminal 10-His tag

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.

N-terminal Sequence Analysis

Ala21

Predicted Molecular Mass

33 kDa

SDS-PAGE

41 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Mca-RPPGFSAFK(Dnp)-OH (Catalog # ES005).
The specific activity is >800 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

934-CY
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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 opening.

Background: Cathepsin X/Z/P

Cathepsin X (also known as Cathepsin Z and P) is a cysteine protease of the papain family (1-5). Compared to other members of the papain family, Cathepsin X has a short proregion and unique insertions. The cysteine residue in the proregion forms a covalent and reversible bond with the active site cysteine residue (6). Acting as a carboxypeptidase, Cathepsin X displays a unique specificity (7-10). It is ubiquitously expressed in human tissues and conserved in other species such as mouse, nematode and echiuran. The nematode enzyme is apparently involved in molting of third stage larvae (11).

References

  1. Nagler, D.K. and R. Menard (1998) FEBS Lett. 434:135.
  2. Santamaria, I. et al. (1998) J. Biol. Chem. 273:16816.
  3. Deussing, J. et al. (2000) Biochim. Biophys. Acta 1491:93.
  4. Pungercar, J. and G. Ivanovski (2000) Pflugers Arch. Eur. J. Physiol. 439:R116.
  5. Pungercar, J. et al. (2000) Pflugers Arch. Eur. J. Physiol. 439:R119.
  6. Sivaraman, J. et al. (2000) J. Mol. Biol. 295:935.
  7. Menard, R. et al. (2001) Biol. Chem. 382:839.
  8. Therrien, C. et al. (2001) Biochemistry 40:2702.
  9. Klemencic, I. et al. (2000) Eur. J. Biochem. 267:5404.
  10. Guncar, G. et al. (2000) Structure Fold Des. 8:305.
  11. Lustigman, S. et al. (1996) J. Biol. Chem. 271:30181.

Alternate Names

Cathepsin P, Cathepsin Z, CTSZ

Entrez Gene IDs

1522 (Human); 64138 (Mouse)

Gene Symbol

CTSZ

UniProt

Q9UBR2

Additional Cathepsin X/Z/P Products

Product Documents for Recombinant Human Cathepsin X/Z/P Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Cathepsin X/Z/P Protein, CF

For research use only

Citations for Recombinant Human Cathepsin X/Z/P Protein, CF

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Protocols

View specific protocols for Recombinant Human Cathepsin X/Z/P Protein, CF (934-CY):

Materials
  • Activation Buffer: 25 mM Sodium Acetate, pH 3.5
  • Assay Buffer: 25 mM Sodium Acetate, 5 mM Dithiothreitol (DTT), pH 3.5
  • Dithiothreitol (DTT), 1 M stock in deionized water
  • Recombinant Human Cathepsin X/Z/P (rhCathepsin X/Z/P) (Catalog # 934-CY)
  • Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Activate rhCathepsin X/Z/P.
    1. Dilute rhCathepsin X/Z/P to 20 µg/mL in Activation Buffer.
    2. Dilute DTT stock to 10 mM in Activation Buffer.
    3. Mix equal volumes of 20 µg/mL rhCathepsin X/Z/P and 10 mM DTT.
    4. Incubate at room temperature for 5 minutes.
  2. Dilute activated rhCathepsin X/Z/P to 0.4 ng/µL in Assay Buffer.
  3. Dilute Substrate to 20 µM in Assay Buffer.
  4. Load 50 µL of the 0.4 ng/µL rhCathepsin X/Z/P in a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 20 µM Substrate without any rhCathepsin X/Z/P.
  5. Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
  6. 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 X/Z/P: 0.02 µg
  • Substrate: 10 µM

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