Cystatin C is a member of family 2 of the Cystatin superfamily (1). It is involved in processes such as tumor invasion and metastasis, inflammation and some neurological diseases. It inhibits many cysteine proteases such as papain and cathepsins B, H, K, L and S (2, 3). It is ubiquitous in human tissues and body fluids. A point mutation in the gene coding for the 120 amino acid mature Cystatin C causes a hereditary form of amyloid angiopathy in which the protein variant (Leu68 to Gln) is deposited in the cerebral arteries, leading to fatal cerebral hemorrhage (4). Cystatin C may have additional clinical applications. For example, it is a good marker for glomerular filtration rate (5).
Recombinant Human Cystatin C Protein, CF
R&D Systems | Catalog # 1196-PI
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Key Product Details
- R&D Systems NS0-derived Recombinant Human Cystatin C Protein (1196-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
Applications
Inhibition Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human Cystatin C protein
Met1-Ala146, with a C-terminal 10-His tag
Met1-Ala146, with a C-terminal 10-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ser27
Predicted Molecular Mass
15 kDa
SDS-PAGE
17 kDa, reducing conditions
Activity
Measured by its ability to inhibit papain cleavage of a fluorogenic peptide substrate Z-FR-AMC (Catalog # ES009).
The IC50 value is <12 nM, under the described conditions.
The IC50 value is <12 nM, under the described conditions.
Reviewed Applications
Read 2 reviews rated 5 using 1196-PI in the following applications:
Formulation, Preparation, and Storage
1196-PI
| Formulation | Lyophilized from a 0.2 μm filtered solution in HEPES and NaCl. |
| Reconstitution | Reconstitute at 100 μg/mL in sterile 25 mM HEPES and 150 mM NaCl, pH 7.8. |
| 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.
|
Calculators
Background: Cystatin C
References
- Reed, C.H. (2000) British J. Biomed. Sci. 57:323.
- Janowski, R. et al. (2001) Nat. Struct. Biol. 8:316.
- Abrahamson, M. (1994) Methods Enzymol. 244:685.
- Abrahamson, M. et al. (1992) Hum. Genet. 89:377.
- Laterza, O.F. et al. (2002) Clin. Chem. 48:699.
Alternate Names
ARMD11, CST3, Gamma-trace, Neuroendocrine basic polypeptide, Post-gamma-globulin
Gene Symbol
CST3
UniProt
Additional Cystatin C Products
Product Documents for Recombinant Human Cystatin C 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.
Product Specific Notices for Recombinant Human Cystatin C Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human Cystatin C Protein, CF
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Protocols
View specific protocols for Recombinant Human Cystatin C Protein, CF (1196-PI):
Materials
- Activation Buffer: 50 mM Tris, 5 mM DTT, pH 7.0
- Assay Buffer: 50 mM Tris, pH 7.0
- Recombinant Human Cystatin C (rhCystatin C) (Catalog # 1196-PI)
- Papain (Sigma, Catalog # P4762)
- Substrate: Z-Phe-Arg-AMC (Catalog # ES009), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Chill Activation Buffer on ice.
- Dilute Papain to 100 µg/mL in Activation Buffer.
- Incubate at room temperature for 15 minutes.
- Prepare a dilution curve of rhCystatin C (MW: 14,709 Da) in Assay Buffer. Make the following serial dilutions: 1000 nM, 500 nM, 333 nM, 222 nM, 111 nM, 55.5 nM, 27.8 nM, 13.9 nM.
- Dilute activated Papain to 2 µg/mL in Assay Buffer.
- Mix equal volumes of the rhCystatin C curve dilutions and the diluted active Papain. Include a Papain control (in duplicate) containing Assay Buffer and the diluted active Papain.
- Incubate mixtures at 37 °C for 10 minutes.
- Dilute Substrate to 200 µM in Assay Buffer.
- Perform a five-fold dilution of the incubated mixture of rhCystatin C and Papain with Assay Buffer.
- Load into a black well plate 50 µL of diluted incubated mixture and start the reaction by adding 50 µL of 200 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm, respectively, for 5 minutes in kinetic mode.
- Derive the 50% inhibiting concentration (IC50) for rhCystatin C by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
- The specific activity for Papain 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 (AMC) (Sigma, Catalog # A-9891).
Per Well:
- Papain: 0.010 µg
- Substrate: 100 µM
- rhCystatin C: 50 nM, 25 nM, 16.65 nM, 11.1 nM, 5.55 nM, 2.78 nM, 1.39 nM, 0.695 nM
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Associated Pathways
