Recombinant Human Leukotriene A4 Hydrolase/LTA4H Protein, CF

R&D Systems | Catalog # 4008-ZN

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Leukotriene A4 Hydrolase/LTA4H Protein (4008-ZN)
  • 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

Sf 21 (baculovirus)

Accession Number

P09960

Applications

Enzyme Activity
View all Leukotriene A4 Hydrolase/LTA4H Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Leukotriene A4 Hydrolase/LTA4H protein
Pro2-Asp611, 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

Pro2

Predicted Molecular Mass

70 kDa

SDS-PAGE

71 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, Arg-7-amido-4-methylcoumarin (R-AMC).
The specific activity is >70 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

4008-ZN
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, and Glycerol.
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: Leukotriene A4 Hydrolase/LTA4H

Leukotrienes are a family of lipid mediators important in a variety of allergic and inflammatory reactions. Synthesized by leukocytes, these molecules are divided into two classes, the spasmogenic cysteinyl leukotrienes and LTB4, a classical chemoattractant (1). Encoded by the LTA4H gene, Leukotriene A4 Hydrolase catalyzes the conversion of unstable epoxide LTA4 to LTB4, which is the final and committed step in LTB4 biosynthesis. As a bifunctional zinc metalloenzyme, LTA4H also acts as an arginyl aminopeptidase (2). LTA4H is a drug target for anti-inflammation, and for cancer prevention and therapy (1, 3).

The mature chain of human LTA4H consists of 610 amino acids (residues 2-611). It is highly specific for LTA4, which also covalently modifies and inhibits the enzyme. The aminopeptidase activity is enhanced by monovalent anions (1). R&D Systems recombinant human LTA4H corresponds to the mature chain, and is characterized by its arginyl aminopeptidase activity.

References

  1. Haeggstrom, J.Z. (2006) J. Biol. Chem. 279:50639.
  2. Orning, L. et al. (1994) J. Biol. Chem. 269:11269.
  3. Chen, X. et al. (2004) Curr. Cancer Drug Targets 4:267.

Alternate Names

LTA4H

Entrez Gene IDs

4048 (Human); 16993 (Mouse); 299732 (Rat)

Gene Symbol

LTA4H

UniProt

P09960

Additional Leukotriene A4 Hydrolase/LTA4H Products

Product Documents for Recombinant Human Leukotriene A4 Hydrolase/LTA4H 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 Leukotriene A4 Hydrolase/LTA4H Protein, CF

For research use only

Citations for Recombinant Human Leukotriene A4 Hydrolase/LTA4H Protein, CF

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Protocols

View specific protocols for Recombinant Human Leukotriene A4 Hydrolase/LTA4H Protein, CF (4008-ZN):

Materials
  • Assay Buffer: 50 mM Tris, 150 mM NaCl, 10 mM CaCl2, pH 7.5 (TCN)
  • Recombinant Human Leukotriene A4 Hydrolase/LTA4H (rhLTA4H) (Catalog # 4008-ZN)
  • Substrate: H-Arg-AMC (Bachem, Catalog # I-1050 or ChemImpex, Catalog # 5859), 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 rhLTA4H to 10 µg/mL in Assay Buffer.
  2. Dilute Substrate to 800 µM in Assay Buffer.
  3. Load 50 µL of 10 µg/mL rhLTA4H into a plate, and start the reaction by adding 50 µL of 800 µM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate.
  4. Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     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-Aldrich, Catalog # A9891).

Per Well:
  • rhLTA4H: 0.5 µg
  • Substrate: 400 µM

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