Recombinant Mouse Cathepsin X/Z/P Protein, CF

Catalog #: 1033-CY
1 Citation Datasheet / COA / SDS

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1033-CY-010

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Mouse Cathepsin X/Z/P 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-RPPGFSAFK(Dnp)-OH (Catalog # ES005). The specific activity is >1,200 pmol/min/µg, as measured under the described conditions.

Source

Mouse myeloma cell line, NS0-derived mouse Cathepsin X/Z/P protein
Ala23-Val306 & Leu27-Val306, both with a C-terminal 10-His tag

Accession #

Q9WUU7

N-terminal Sequence
Analysis

Ala23 & Leu27

Structure / Form

Pro form

Predicted Molecular Mass

33 kDa

SDS-PAGE

40 kDa, reducing conditions

Product Datasheets

1033-CY

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.

1033-CY

Formulation	Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
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 opening.

Assay Procedure

Materials

Assay Buffer: 25 mM NaOAc, 5 mM DTT, pH 3.0
Recombinant Mouse Cathepsin X/Z/P (rmCathepsin X/Z/P) (Catalog # 1033-CY)
Fluorogenic Peptide Substrate V: 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

Dilute rmCathepsin X/Z/P to 10 µg/mL in Assay Buffer.
Incubate at room temperature for 5 minutes to activate.
Dilute activated rmCathepsin X/Z/P to 0.4 ng/µL in Assay Buffer.
Dilute Substrate to 20 µM in Assay Buffer.
Load 50 µL of the 0.4 ng/µL rmCathepsin X/Z/P into a black well plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate without any rmCathepsin X/Z/P.
Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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:

rmCathepsin X/Z/P: 0.02 µg
Substrate: 10 µM

Reconstitution Calculator

Background: Cathepsin X/Z/P

Cathepsin X (also known as Cathepsin Z and P) is a relatively new 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

Deussing, et al. (2000) Biochim. Biophys. Acta 1491:93.
Santamaria, et al. (1998) J. Biol. Chem. 273:16816.
Nagler and Menard (1998) FEBS Lett. 434:135.
Pungercar and Ivanovski (2000) Pflugers Arch. 439:R116.
Pungercar, et al. (2000) Pflugers Arch. 439:R119.
Sivaraman, et al. (2000) J. Mol. Biol. 295:935.
Menard, et al. (2001) Biol. Chem. 382:839.
Therrien, et al. (2001) Biochemistry 40:2702.
Klemencic, et al. (2000) Eur. J. Biochem. 267:5404.
Guncar, et al. (2000) Structure Fold Des. 8:305.
Lustigman, et al. (1996) J. Biol. Chem. 271:30181.

Entrez Gene IDs

1522 (Human); 64138 (Mouse)

Alternate Names

Cathepsin P; Cathepsin X/Z/P; Cathepsin Z; CTSX; CTSZ

Related Research Areas

Citation for Recombinant Mouse Cathepsin X/Z/P 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.

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