Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF

R&D Systems | Catalog # 6069-AO

This item is in process of being discontinued.
R&D Systems
Discontinued Product
6069-AO has been discontinued. View all Lysyl Oxidase Homolog 3/LOXL3 products.

Key Product Details

Source

CHO

Accession Number

P58215

Structure / Form

Monomer

Applications

Enzyme Activity
View all Lysyl Oxidase Homolog 3/LOXL3 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Lysyl Oxidase Homolog 3/LOXL3 protein
Met1-Ile753, with a C-terminal 10-His tag

Purity

>90%, 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

Ser26

Predicted Molecular Mass

82 kDa

SDS-PAGE

90-100 kDa, reducing conditions

Activity

Measured by its ability to produce hydrogen peroxide during the oxidation of benzylamine.
The specific activity is >5 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6069-AO
Formulation Supplied as a 0.2 μm filtered solution in MES 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: Lysyl Oxidase Homolog 3/LOXL3

Lysyl oxidase-like protein 3 (LOXL3) is a secreted copper amine oxidase. The N-terminal region of LOXL3 contains four scavenger receptor cysteine-rich (SRCR) domains. The C-terminal region consists of a catalytic domain similar to other lysyl oxidases and a cytokine receptor-like domain (1). The catalytic domain contains conserved residues required for copper binding and formation of a lysyl tyrosylquinone cofactor (2). LOXL3 is expressed primarily in epidermis, kidney, aorta, and chondrocytes (3). The enzyme is expressed at low levels in most other tissues. Like lysyl oxidase, LOXL3 oxidizes the lysine epsilon -amino groups of a number of collagen substrates, and is also active against other primary amines (4). However, LOXL3 does not require proteolytic processing to be activated.

References

  1. Csiszar, K. (2001) Prog. Nucl. Acid Res. Mol. Biol. 70:1.
  2. Maki, J.M. and K.I. Kivirikko (2001) Biochem J. 355:381.
  3. Molnar, J. et al. (2003) Biochim. Biophys. Acta 1647:220.
  4. Lee, J. and Y. Kim (2006) J. Biol. Chem. 281:37282.

Alternate Names

LOL3, LOXL3

Entrez Gene IDs

84695 (Human); 16950 (Mouse)

Gene Symbol

LOXL3

UniProt

P58215

Additional Lysyl Oxidase Homolog 3/LOXL3 Products

Product Documents for Recombinant Human Lysyl Oxidase Homolog 3 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.

Download SDS

Product Specific Notices for Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF

For research use only

Related Research Areas

Oxidases and Oxygenases
Redox Enzymes

Citations for Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF

Customer Reviews for Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF and earn rewards!

Have you used Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Lysyl Oxidase Homolog 3 Protein, CF (6069-AO):

Materials
  • Assay Buffer: 50 mM Sodium Borate, 250 mM Urea, 10 mM CaCl2, pH 8.0 
  • Recombinant Human Lysyl Oxidase Homolog 3/LOXL3 (rhLOXL3) (Catalog # 6069-AO)
  • Coupling Enzyme: Horseradish Peroxidase (HRP) (250-330 U/mg) (Sigma, Catalog # P8375), 250 units/mL stock in 0.1 M Sodium Phosphate, pH 8.0
  • Substrate Component 1: Benzylamine (Sigma, Catalog # B5136), 100 mM stock in deionized water
  • Substrate Component 2: Amplex Ultra Red (AUR) (Molecular Probes, Catalog # A36006), 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 rhLOXL3 to 40 ng/µL in Assay Buffer.
  2. Dilute Benzylamine to 16 mM in Assay Buffer.
  3. Combine equal volumes of 40 ng/µL rhLOXL3 and 16 mM Benzylamine. Also create a Substrate Blank by combining equal volumes of Assay Buffer and 16 mM Benzylamine.
  4. Incubate the reactions for 30 minutes at 37 °C.
  5. Prepare the Substrate Mixture containing 2 units/mL HRP and 40 µM AUR in Assay Buffer.
  6. Load 50 µL of the incubated reactions into the wells of a black well plate, and add 50 µL of Substrate Mixture to each well.
  7. Read at excitation and emission wavelengths of 544 nm and 590 nm (top read), respectively, in endpoint mode. Note: A cutoff must be set manually at a wavelength of 570 nm. 
  8. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

*Adjusted for Substrate Blank.
**Derived using a fluorescent standard prepared by incubating 20 µM AUR, 1 unit/mL HRP, 4 mM Benzylamine, and a curve of Hydrogen Peroxide (Sigma, Catalog # H1009) in Assay Buffer. Use this oxidized AUR curve to determine the conversion factor.

Per Well:
  • rhLOXL3: 1.0 µg
  • Benzylamine: 4 mM
  • HRP: 1 unit/mL
  • AUR: 20 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes