Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF

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1 μg/lane of Recombinant Human Lysyl Oxidase Homolog 2 (Catalog # 2639-AO) and 1 μg/lane of competitor Lysyl Oxidase Homolog 2 were resolved with 4-20% SDS-PAGE under reducing (R) and non-reducing (NR) conditions and ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF Summary

Details of Functionality
Measured by its ability to produce hydrogen peroxide during the oxidation of benzylamine. The specific activity is >2 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Lysyl Oxidase Homolog 2/LOXL2 protein
Met1-Gln774, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Low recovery, Gln26 predicted
Structure / Form
Monomer
Protein/Peptide Type
Recombinant Enzymes
Gene
LOXL2
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
85 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
100-110 kDa, reducing conditions
Publications
Read Publications using
2639-AO 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in MES and NaCl.
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Sodium Borate, 1.2 M Urea, 10 mM CaCl2, pH 8.0 
  • Recombinant Human Lysyl Oxidase Homolog 2/LOXL2 (rhLOXL2) (Catalog # 2639-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 rhLOXL2 to 10 ng/µL in Assay Buffer.
  2. Dilute Benzylamine to 4 mM in Assay Buffer.
  3. Combine equal volumes of 10 ng/µL rhLOXL2 and 4 mM Benzylamine. Also create a Substrate Blank by combining equal volumes of Assay Buffer and 4 mM Benzylamine.
  4. Incubate the reactions for 30 minutes at 37 °C.
  5. Prepare the Substrate Mixture, 2 units/mL HRP and 40 µM AUR, in Assay Buffer.
  6. Load 50 µL of the incubated reactions into the wells of a plate and add 50 µL of Substrate Mixture.
  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, 1 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:
  • rhLOXL2: 0.25 µg
  • Benzylamine: 1 mM
  • HRP: 1 unit/mL
  • AUR: 20 µM

Notes

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

Alternate Names for Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF

  • EC 1.4.3
  • EC 1.4.3.-
  • LOL2
  • LOR2
  • LOXL2
  • Lysyl Oxidase Homolog 2
  • lysyl oxidase related 2
  • lysyl oxidase-like 2
  • Lysyl oxidase-like protein 2
  • Lysyl oxidase-related protein 2
  • Lysyl oxidase-related protein WS9-14
  • WS9-14

Background

Lysyl Oxidase Homolog 2 (lysyl oxidase-like protein 2, LOXL2) is a member of lysyl oxidase-like (LOXL) gene family which includes LOXL1 through LOXL4. These enzymes are secreted copper-binding amine oxidases that oxidize primary amine substrates to aldehydes (1). The N-terminal region of LOXL2 contains four scavenger receptor cysteine-rich (SRCR) domains, and the C-terminal region is a catalytic domain similar to other lysyl oxidases (1). The catalytic domain contains conserved residues required for copper binding and formation of a lysyl tyrosylquinone co-factor (2). Although some of the LOXL enzymes are known to cross-link collagen and elastin substrates, such a function has yet to be characterized for LOXL2. It has been shown that LOXL2 promotes cell migration and tumor cell invasiveness (3, 4). Elevated expression of LOXL2 is also associated with cancer progression in various tumors and carcinoma cell lines, which makes it a potential marker for prognosis of cancer (5). LOXL2 is expressed in many tissues, with elevated levels in reproductive tissues such as placenta, uterus, and prostate (6).
  1. Csiszar, H. (2001) Prog. Nucleic Acid Res. Mol. Biol. 70:1.
  2. Maki, J.M. and K.I. Kivirikko (2001) Biochem J. 355:381.
  3. Akiri, G. et al. (2003) Cancer Res. 63:1657.
  4. Hollosi, P. et al. (2009) Int. J. Cancer. 125:318.
  5. Peinado, H. et al. (2008) Cancer Res. 68:4541.
  6. Jourdan-Le Saux C. et al. (1999) J. Biol. Chem. 274:12939.

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2639-AO
Species: Hu
Applications: Enzyme Activity

Publications for Lysyl Oxidase Homolog 2/LOXL2 (2639-AO)(6)

We have publications tested in 3 confirmed species: Human, Mouse, NA.

We have publications tested in 2 applications: Bioassay, Enzyme Assay.


Filter By Application
Bioassay
(3)
Enzyme Assay
(3)
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Human
(4)
Mouse
(1)
NA
(1)
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Showing Publications 1 - 6 of 6.
Publications using 2639-AO Applications Species
DJ Leeming, N Willumsen, JMB Sand, S Holm Niels, B Dasgupta, C Brodmerkel, M Curran, CL Bager, MA Karsdal A serological marker of the N-terminal neoepitope generated during LOXL2 maturation is elevated in patients with cancer or idiopathic pulmonary fibrosis Biochem Biophys Rep, 2019;17(0):38-43. 2019 [PMID: 30555938] (Bioassay, Human) Bioassay Human
Schilter H, Findlay A, Perryman L, Yow T, Moses J, Zahoor A, Turner C, Deodhar M, Foot J, Zhou W, Greco A, Joshi A, Rayner B, Townsend S, Buson A, Jarolimek W The lysyl oxidase like 2/3 enzymatic inhibitor, PXS-5153A, reduces crosslinks and ameliorates fibrosis. J Cell Mol Med, 0;0(0):. 0 [PMID: 30536539] (Bioassay, NA) Bioassay NA
Y Wei, TJ Kim, DH Peng, D Duan, DL Gibbons, M Yamauchi, JR Jackson, CJ Le Saux, C Calhoun, J Peters, R Derynck, BJ Backes, HA Chapman Fibroblast-specific inhibition of TGF-?1 signaling attenuates lung and tumor fibrosis J. Clin. Invest., 2017;0(0):. 2017 [PMID: 28872461] (Bioassay, Mouse) Bioassay Mouse
V Aumiller, B Strobel, M Romeike, M Schuler, BE Stierstorf, S Kreuz Comparative analysis of lysyl oxidase (like) family members in pulmonary fibrosis Sci Rep, 2017;7(1):149. 2017 [PMID: 28273952] (Enzyme Assay, Human) Enzyme Assay Human
Schilter H, Collison A, Russo R, Foot J, Yow T, Vieira A, Tavares L, Mattes J, Teixeira M, Jarolimek W Effects of an anti-inflammatory VAP-1/SSAO inhibitor, PXS-4728A, on pulmonary neutrophil migration. Respir Res, 2015;16(0):42. 2015 [PMID: 25889951] (Enzyme Assay, Human) Enzyme Assay Human
Vonk WI, de Bie P, Wichers CG, van den Berghe PV, van der Plaats R, Berger R, Wijmenga C, Klomp LW, van de Sluis B The copper-transporting capacity of ATP7A mutants associated with Menkes disease is ameliorated by COMMD1 as a result of improved protein expression. Cell. Mol. Life Sci., 2012;69(1):149-63. 2012 [PMID: 21667063] (Enzyme Assay, Human) Enzyme Assay Human

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Bioinformatics

Gene Symbol LOXL2
Uniprot