Recombinant Human Cystatin F Protein, CF

Images

 
There are currently no images for Cystatin F (1889-PI).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Inhibition Activity
Format
Carrier-Free

Order Details

Recombinant Human Cystatin F Protein, CF Summary

Details of Functionality
Measured by its ability to inhibit active Cathepsin L cleavage of a fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008). The IC50 value is <6.0 nM, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Cystatin F protein
Gly20-His145, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Gly20
Structure / Form
Disulfide-linked homodimer
Protein/Peptide Type
Recombinant Enzymes
Gene
CST7
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
15 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
16 kDa, 21 kDa and 26 kDa, reducing conditions
Publications
Read Publications using
1889-PI in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Lyophilized from a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile 25 mM Tris and 150 mM NaCl, pH 7.5.
Assay Procedure
  • Assay Buffer: 50 mM MES, pH 6.0
  • Dithiothreitol (DTT) (Sigma, Catalog # D0632), 1 M stock in deionized water
  • Recombinant Human Cystatin F (rhCystatin F) (Catalog # 1889-PI)
  • Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
  • Substrate: Z-Leu-Arg-AMC (Catalog # ES008), 10 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCathepsin L to 1 µg/mL in Assay Buffer with 5 mM DTT.
  2. Incubate on ice for 15 minutes.
  3. After incubation, dilute activated rhCathepsin L to 0.5 µg/mL in Assay Buffer.
  4. Prepare a curve of rhCystatin F (MW: 15,356 Da) in Assay Buffer. Make the following serial dilutions: 2000, 1000, 500, 250, 125, 62.5, 31.3, 15.6, 7.8, and 3.9 nM.
  5. Gently mix equal volumes of the rhCystatin F 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 room temperature for 15 minutes.
  7. Make a five-fold dilution of reaction mixture with Assay Buffer.
  8. Dilute Substrate to 20 µM in Assay Buffer.
  9. Load 50 µL of the incubated mixtures into a black well plate, 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 50% inhibition concentration (IC50) value for rhCystatin F by plotting RFU/min (or specific activity) versus 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 F curve: 100, 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78, 0.39 and 0.20 nM
  • Substrate: 10 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Cystatin F Protein, CF

  • CMAP
  • CST7
  • Cystatin 7
  • cystatin F (leukocystatin)
  • Cystatin F
  • Cystatin-7
  • cystatin-F
  • Cystatin-like metastasis-associated protein
  • Leukocystatin

Background

Cystatin F, also known as leukocystatin and CMAP (Cystatin‑like Metastasis‑Associated Protein), is a member of the cystatin superfamily (1‑3). Cystatin F is selectively expressed by hematopoietic cells and may be a biomarker for both liver metastasis and inflammatory lung disorders (3, 4). As a cysteine protease inhibitor, it shows selectivity towards cathepsin L and legumain (1, 2, 5). Compared to other secreted cystatins including C, D, E/M, S, SA and SN, which contain two intra disulfide bonds, cystatin F has two extra Cys residues that may be involved in inter disulfide bonds. Indeed, rhCystatin F showed disulfide bond‑linked dimer formation, which was also the case for an E. coli expressed fusion protein containing mature human cystatin F and glutathione S‑transferase (2).

  1. Ni, J. et al. (1998) J. Biol. Chem. 273:24797.
  2. Halfon, S. et al. (1998) J. Biol. Chem. 273:16400.
  3. Utsunomiya, T. et al. (2002) Clin Cancer 8:2591
  4. Werle, B. et al. (2003) Biol. Chem. 384:281.
  5. Gruninger-Leitch, F. et al. (2000) Nat. Biotechnol. 18:66.

Customers Who Viewed This Item Also Viewed...

NBP1-88769
Species: Hu, Mu
Applications: WB, IHC, IHC-P
NBP2-24915
Species: Hu, Mu, Rt, Bv
Applications: WB, IHC-P
AF1436
Species: Mu
Applications: WB
AF6108
Species: Hu
Applications: IHC
AF952
Species: Hu
Applications: WB, IHC, IP, ELISA(Cap), ELISA(Det), ELISA(Sta)
AF3814
Species: Hu, Mu, Rt
Applications: WB, IP
NBP2-34390
Species: Hu
Applications: WB, Flow, ICC/IF, IHC-Fr, IHC-P, IP
NBP1-78977
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC/IF, IHC, IHC-P
NBP2-52575
Species: Hu
Applications: WB, ELISA, Flow, IHC, IHC-P, CyTOF-ready, ICC, IF
NBP2-41211
Species: Hu, Mu, Rt
Applications: WB, ELISA, IHC, IHC-P, IF
NB110-59086
Species: Hu, Pm, Bv(-), Mu(-), Rt(-)
Applications: WB, IHC, IHC-P
NBP2-22449
Species: Hu, Mu, Rt, Fe, GP, Rb
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-P, IP, GS
AF745
Species: Mu
Applications: WB, Block
AF1286
Species: Hu
Applications: WB, Simple Western, IHC, IP
AF2340
Species: Hu
Applications: WB, Flow, IP, CyTOF-ready
NBP2-56762
Species: Hu
Applications: WB, ICC/IF
NBP2-20383
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P

Publications for Cystatin F (1889-PI)(2)

Reviews for Cystatin F (1889-PI) (0)

There are no reviews for Cystatin F (1889-PI). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen
  • 1st to review with an image -- $50/€35/£30/$50 CAD/¥300 Yuan/¥5000 Yen

FAQs for Cystatin F (1889-PI) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Additional Array Products

Bioinformatics Tool for Cystatin F (1889-PI)

Discover related pathways, diseases and genes to Cystatin F (1889-PI). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for Cystatin F (1889-PI)

Discover more about diseases related to Cystatin F (1889-PI).
 

Pathways for Cystatin F (1889-PI)

View related products by pathway.

PTMs for Cystatin F (1889-PI)

Learn more about PTMs related to Cystatin F (1889-PI).

Blogs on Cystatin F

There are no specific blogs for Cystatin F, but you can read our latest blog posts.

Customers Who Bought This Also Bought

GM-CSF [Unconjugated]
215-GM-010/CF

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant Human Cystatin F Protein, CF and receive a gift card or discount.

Bioinformatics

Gene Symbol CST7
Entrez
Uniprot