Recombinant Human Testican 1/SPOCK1 Protein, CF

Images

 
There are currently no images for Testican 1/SPOCK1 (2327-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 Testican 1/SPOCK1 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 <10 nM, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Testican 1/SPOCK1 protein
Arg22-Trp439, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Arg22
Protein/Peptide Type
Recombinant Proteins
Gene
SPOCK1
Purity
>90%, 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
48 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
50-56 kDa, reducing conditions
Publications
Read Publication using
2327-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 PBS.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 μg/mL in sterile PBS.
Assay Procedure
  • Assay Buffer: 50 mM MES, 5 mM DTT, pH 6.0
  • Recombinant Human Testican 1/SPOCK1 (rhTestican 1) (Catalog # 2327-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 1 µg/mL in Assay Buffer.
  2. Incubate diluted rhCathepsin L on ice for 15 minutes.
  3. Dilute incubated rhCathepsin L to 0.25 µg/mL in Assay Buffer.
  4. Prepare a curve of rhTestican 1 (MW: 47,800 Da) in Assay Buffer. Make the following serial dilutions: 500, 250, 125, 83.3, 55.6, 37, 24.7, and 12.3 nM.
  5. Combine equal volumes of diluted rhCathepsin L and rhTestican 1 at each concentration of the curve. Include two controls containing equal volumes of Assay buffer and diluted rhCathepsin L without any rhTestican 1.
    Incubate mixtures at 37 ˚C for 15 minutes.
  6. Dilute the reaction mixtures 1/5 in Assay Buffer.
  7. Dilute Substrate to 20 µM in Assay Buffer.
  8. Load 50 µL of the incubated mixtures in a plate, and start the reaction by adding 50 µL of 20 µM Substrate.
  9. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  10. Derive the 50% inhibition concentration (IC50) value for rhTestican 1 by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
  11. 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 (AMC) (Sigma, Catalog # A-9891).

Per Well:
  • rhCathepsin L: 0.00125 µg
  • rhTestican 1 curve: 25, 12.5, 6.25, 4.165, 2.78, 1.85, 1.235, and 0.615 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 Testican 1/SPOCK1 Protein, CF

  • FLJ37170
  • Protein SPOCK
  • sparc/osteonectin, cwcv and kazal-like domains proteoglycan (testican) 1
  • SPOCK1
  • SPOCKcwcv and kazal-like domains proteoglycan (testican)
  • Testican 1
  • TIC1
  • TICN1

Background

Testican 1, encoded by the SPOCK1 gene, is a proteoglycan first identified in human seminal plasma (1). The cDNAs from the human testis and mouse brain indicate 95% identity between the deduced amino acid sequences from the two species, predicting a conserved function (2, 3). Human Testican 1 is able to inhibit attachment of Neuro-2a cells and their ability to form neurite extensions (4). At R&D Systems, recombinant human (rh) Testican 1 has also been shown to be able to enhance neurite outgrowth of E18 rat embryonic hippocampal neurons. Testican 1 contains Ca2+-binding domain and the C-terminal acidic domain with putative glycosaminoglycan attachment sites (5). In addition, it contains three potential inhibitory domains targeted toward three different classes of proteases, metallo, cysteine and serine proteases. The N-terminal region, which is unique to testicans, is responsible for the inhibition of testican 1 towards MMP-14 (MT1-MMP, a metalloprotease) activation of MMP-2 (6). The thyropin domain may be responsible for the inhibition of testican 1 towards cathepsin L, a cysteine protease (5). The follistatin-like domain with a six cysteine Kazal-like motif may inhibit serine proteases. The purified rhTestican 1 is capable of inhibiting rhMMP-14 and rhCathepsin L (Catalog # 918-MP and 952-CY) in assays using the fluorogenic peptide substrates (Catalog # ES001 and ES008).

  1. Bonnet, F. et al. (1992) Biochem. J. 288:565.
  2. Alliel, P.M. et al. (1993) Eur. J. Biochem. 214:347.
  3. Bonnet, F. et al. (1996) J. Biol. Chem. 271:565.
  4. Marr, H.S. and C.-J.S Edgell (2003) Matrix Biol. 22:259.
  5. Bocock, J.P. et al. (2003) Eur. J. Biochem. 270:4008.
  6. Nakada, M. et al. (2001) Cancer Res. 61:8896.

Customers Who Viewed This Item Also Viewed...

NBP1-80972
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P
NBP1-92442
Species: Hu
Applications: WB, IHC, IHC-P
AF952
Species: Hu
Applications: WB, IHC, IP, ELISA(Cap), ELISA(Det), ELISA(Sta)
NB200-193
Species: Hu, Mu, Rt, Po, Bv, Ch, Eq
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
AF2728
Species: Hu
Applications: WB, IHC, CyTOF-ready, ICFlow
NBP2-52918
Species: Hu
Applications: WB, ICC/IF, IHC, IHC-P, KD
AF2346
Species: Mu
Applications: WB, IP
NBP2-67415
Species: Hu, Mu, Rt
Applications: WB, Flow, ICC/IF, IHC, IHC-P, IP
H00006941-M01
Species: Hu
Applications: WB, ELISA, ICC/IF, IHC-P, RNAi
AF808
Species: Mu
Applications: WB, IHC, ELISA(Cap), ELISA(Det), Neut, ELISA(Sta)
NBP1-90312
Species: Hu
Applications: IHC, IHC-P
NBP1-88227
Species: Hu
Applications: WB, IHC, IHC-P
NBP2-34294
Species: Hu, Mu, Rt
Applications: WB, Flow, IHC, IHC-P
AF1785
Species: Hu
Applications: WB, IHC, IP
AF2818
Species: Hu
Applications: WB, ICC
NBP1-80918
Species: Hu
Applications: IHC, IHC-P
8115-SA
Species: Hu
NB100-64980
Species: Hu
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
2327-PI
Species: Hu
Applications: Inhibition Activity

Publications for Testican 1/SPOCK1 (2327-PI)(1)

We have publications tested in 1 confirmed species: N/A.

We have publications tested in 1 application: WB Ctrl.


Filter By Application
WB Ctrl
(1)
All Applications
Filter By Species
N/A
(1)
All Species

Reviews for Testican 1/SPOCK1 (2327-PI) (0)

There are no reviews for Testican 1/SPOCK1 (2327-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 Testican 1/SPOCK1 (2327-PI) (0)

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

Additional Array Products

Bioinformatics Tool for Testican 1/SPOCK1 (2327-PI)

Discover related pathways, diseases and genes to Testican 1/SPOCK1 (2327-PI). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for Testican 1/SPOCK1 (2327-PI)

Discover more about diseases related to Testican 1/SPOCK1 (2327-PI).
 

Pathways for Testican 1/SPOCK1 (2327-PI)

View related products by pathway.

PTMs for Testican 1/SPOCK1 (2327-PI)

Learn more about PTMs related to Testican 1/SPOCK1 (2327-PI).

Blogs on Testican 1/SPOCK1

There are no specific blogs for Testican 1/SPOCK1, but you can read our latest blog posts.

Customers Who Bought This Also Bought

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 Testican 1/SPOCK1 Protein, CF and receive a gift card or discount.

Bioinformatics

Gene Symbol SPOCK1
Uniprot