Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF

Catalog # Availability Size / Price Qty
2639-AO-010
Recombinant Human Lysyl Oxidase Homolog 2 Protein SDS-PAGE
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Product Details
Citations (6)
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Reviews (1)

Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF Summary

Product Specifications

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.
Activity
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
Analysis
Low recovery, Gln26 predicted
Structure / Form
Monomer
Predicted Molecular Mass
85 kDa
SDS-PAGE
100-110 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

2639-AO

Formulation Supplied as a 0.2 μm filtered solution in MES and NaCl.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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

Data Image

SDS-PAGE Recombinant Human Lysyl Oxidase Homolog 2 Protein SDS-PAGE View Larger

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 visualized by silver staining.

Reconstitution Calculator

Reconstitution Calculator

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Background: Lysyl Oxidase Homolog 2/LOXL2

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).

References
  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.
Entrez Gene IDs
4017 (Human); 94352 (Mouse); 102138804 (Cynomolgus Monkey)
Alternate Names
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

Citations for Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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  1. A serological marker of the N-terminal neoepitope generated during LOXL2 maturation is elevated in patients with cancer or idiopathic pulmonary fibrosis
    Authors: DJ Leeming, N Willumsen, JMB Sand, S Holm Niels, B Dasgupta, C Brodmerkel, M Curran, CL Bager, MA Karsdal
    Biochem Biophys Rep, 2019;17(0):38-43.
    Species: Human
    Sample Types: Protein
    Applications: Bioassay
  2. Comparative analysis of lysyl oxidase (like) family members in pulmonary fibrosis
    Authors: V Aumiller, B Strobel, M Romeike, M Schuler, BE Stierstorf, S Kreuz
    Sci Rep, 2017;7(1):149.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  3. Fibroblast-specific inhibition of TGF-?1 signaling attenuates lung and tumor fibrosis
    Authors: 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
    J. Clin. Invest., 2017;0(0):.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Effects of an anti-inflammatory VAP-1/SSAO inhibitor, PXS-4728A, on pulmonary neutrophil migration.
    Authors: Schilter H, Collison A, Russo R, Foot J, Yow T, Vieira A, Tavares L, Mattes J, Teixeira M, Jarolimek W
    Respir Res, 2015;16(0):42.
    Species: Human
    Sample Types: Protein
    Applications: Enzyme Assay
  5. The copper-transporting capacity of ATP7A mutants associated with Menkes disease is ameliorated by COMMD1 as a result of improved protein expression.
    Authors: 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
    Cell. Mol. Life Sci., 2012;69(1):149-63.
    Species: Human
    Sample Types: Whole Cells
    Applications: Enzyme Assay
  6. The lysyl oxidase like 2/3 enzymatic inhibitor, PXS-5153A, reduces crosslinks and ameliorates fibrosis.
    Authors: 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
    J Cell Mol Med, 0;0(0):.
    Species: NA
    Sample Types: Collagen
    Applications: Bioassay

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Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF
By Anonymous on 08/18/2017
Application: Binding assay/Protein-protein interaction