Recombinant Human Lipocalin-1 Protein, CF

R&D Systems | Catalog # 1708-PI

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Lipocalin-1 Protein (1708-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

P31025

Applications

Inhibition Activity
View all Lipocalin-1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Lipocalin-1 protein
His19-Asp176, 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

His19

Predicted Molecular Mass

19 kDa

SDS-PAGE

20-22 kDa, reducing conditions

Activity

Measured by its ability to inhibit active Cathepsin V cleavage of a fluorogenic peptide substrate Z-LR-AMC (Catalog # ES008).
The IC50 value is <400 nM, as measured under the described conditions.

Formulation, Preparation, and Storage

1708-PI
Formulation Supplied as a 0.2 μm filtered solution in Tris 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.

Background: Lipocalin-1

Lipocalin-1, also known as tear prealbumin or von Ebner’s gland protein (VEGP), is encoded by the LCN1 gene (1-3). It is a member of the Lipocalin superfamily that binds many different classes of lipophylic chemicals (4). Lipocalin-1 contains three sequence motifs similar to the cystatins, a superfamily of cysteine protease inhibitors (5). In fact, it has been suggested that Lipocalin-1 is a physiological inhibitor of cysteine proteases and plays a role in the control of inflammatory processes in oral and ocular tissues (5). Recombinant Human Lipocalin-1 corresponds to the mature and secreted protein. It is a weak inhibitor of cysteine proteases such as cathepsin V, which is similar to recombinant human Cystatin S (Catalog # http://www.rndsystems.com/product_results.aspx?k=1296-PI">1296-PI).

References

  1. Redl, B. et al. (1992) J. Biol. Chem. 267:20282.
  2. Blaker, M. et al. (1993) Biochim. Biophys. Acta 1172:131.
  3. Lassagne, H. and A.M. Gachon (1993) Exp. Eye Res. 56:605.
  4. Redl, B. et al. (2000) Biochim. Biophys. Acta 1482:241.
  5. van’t Hof, W. et al. (1997) J. Biol. Chem. 272:1837.

Alternate Names

LCN1, Lipocalin1, Tear Prealbumin, VEGP

Entrez Gene IDs

3933 (Human)

Gene Symbol

LCN1

UniProt

P31025

Additional Lipocalin-1 Products

Product Documents for Recombinant Human Lipocalin-1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Lipocalin-1 Protein, CF

For research use only

Citations for Recombinant Human Lipocalin-1 Protein, CF

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Protocols

View specific protocols for Recombinant Human Lipocalin-1 Protein, CF (1708-PI):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, pH 5.5
  • Cathepsin Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, 10 mM DTT, pH 5.5
  • Substrate Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, 5 mM DTT, pH 5.5
  • Recombinant Human Lipocalin-1 (rhLipocalin-1) (Catalog # 1708-PI)
  • Recombinant Human Cathepsin V (rhCathepsin V) (Catalog # 1080-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. Prepare a curve of rhLipocalin-1 (MW: 18,805 Da) in Assay Buffer. Make the following serial dilutions:  40,000, 10,000, 4000, 1000, 500, 250, 125, 50, 12.5, and 6.25 nM.
  2. Dilute rhCathepsin V to 2 µg/mL in Cathepsin Buffer.
  3. Mix equal volumes of the rhLipocalin-1 curve dilutions and the 2 µg/mL rhCathepsin V. Include a Cathepsin Control (in duplicate) containing Assay Buffer in place of rhLipocalin-1.
  4. Incubate mixtures at room temperature for 30 minutes.
  5. Dilute Substrate to 40 µM in Substrate Buffer.
  6. Load into a black well plate 50 µL of the incubated mixtures, and start the reaction by adding 50 µL of 40 µM Substrate.
  7. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  8. Derive the 50 % inhibiting concentration (IC50) of rhLipocalin-1 by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
  9. The specific activity for rhCathepsin V 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 V: 0.05 µg
  • rhLipocalin-1: 10,000, 2500, 1000, 250, 125, 62.5, 31.25, 12.5, 3.125, and 1.5625 nM
  • Substrate: 20 µM

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