Recombinant Human Cystatin SN Protein, CF

R&D Systems | Catalog # 1285-PI

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Human Cystatin SN Protein (1285-PI)
  • 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

P01037

Applications

Inhibition Activity
View all Cystatin SN Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Cystatin SN protein
Trp21-Ser141, 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

Trp21

Predicted Molecular Mass

16 kDa

SDS-PAGE

16 kDa, reducing conditions

Activity

Measured by its ability to inhibit active Cathepsin L cleavage of a fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008).
The IC50 value is <7.0 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1285-PI
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.

Calculators

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.

=
÷

Background: Cystatin SN

Cystatin SN is a member of family 2 of the cystatin superfamily (1). Together with cystatins S and SA, it is produced by the salivary gland and secreted largely in the submandibular/sublingual saliva (2). Cystatin SN inhibits members of the papain family including cathepsins B, C, H and L (3).

References

  1. Abrahamson, M. (1994) Methods Enzymol. 244:685.
  2. Baron, A.C. et al. (1999) Oral Dis. 5:344.
  3. Baron, A. et al. (1999) Oral Dis. 5:234.

Alternate Names

CST1

Entrez Gene IDs

1469 (Human)

Gene Symbol

CST1

UniProt

P01037

Additional Cystatin SN Products

Product Documents for Recombinant Human Cystatin SN Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Cystatin SN Protein, CF

For research use only

Related Research Areas

Cysteine Proteases and Regulators

Citations for Recombinant Human Cystatin SN Protein, CF

Customer Reviews for Recombinant Human Cystatin SN Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Cystatin SN Protein, CF and earn rewards!

Have you used Recombinant Human Cystatin SN Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Cystatin SN Protein, CF (1285-PI):

Materials
  • Activation Buffer: 50 mM MES, 5 mM DTT, pH 6.0
  • Assay Buffer: 50 mM MES, pH 6.0
  • Recombinant Human Cystatin SN (rhCystatin SN) (Catalog # 1285-PI)
  • Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
  • Substrate: Z-Leu-Arg-AMC (Catalog # ES008)
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin L to 40 µg/mL in Activation Buffer.
  2. Incubate 15 minutes on ice.
  3. Dilute rhCathepsin L to 0.5 µg/mL in Activation Buffer.
  4. Prepare a curve of rhCystatin SN (MW: 15,677 Da) in Assay Buffer. Make the following serial dilutions: 2000, 500, 200, 100, 66.7, 44.4, 22.2, 11.1, and 2.78 nM.
  5. Gently mix equal volumes of the rhCystatin SN curve dilutions and the diluted active rhCathepsin L. Include a control (in duplicate) containing Assay Buffer and the diluted active rhCathepsin L.
  6. Incubate mixtures at 37 ºC for 15 minutes.
  7. After incubation, perform a 1/5 dilution of each reaction mixture with Assay Buffer.
  8. Dilute Substrate to 20 µM in Assay Buffer.
  9. Load into a black well plate 50 µL of the incubated mixtures, and start the reaction by adding 50 µL of 20 µM Substrate.
  10. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  11. Derive the 50% inhibition concentration (IC50) value for rhCystatin SN by plotting RFU/min (or specific activity) vs. concentration with 4-PL fitting.
  12. The specific activity for rhCathepsin L at each point may be determined using the following formula (if needed):

     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 7-amino, 4-Methyl Coumarin (Sigma, Catalog # A-9891).

Per Well:

  • rhCathepsin L: 0.0025 µg
  • rhCystatin SN curve: 100, 25, 10, 5, 3.33, 2.22, 1.11, 0.555, and 0.139 nM
  • Substrate: 10 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes