Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF
Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF Summary
Product Specifications
Met1-Gln774, with a C-terminal 10-His tag
Analysis
Product Datasheets
Carrier Free
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.
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.
|
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 units/mL stock in 0.1 M Sodium Phosphate, pH 8.0
- Substrate Component 1: Benzylamine, 100 mM stock in deionized water
- Substrate Component 2: Amplex Ultra Red (AUR), 10 mM stock in DMSO
- 96-well Black Plate
- Plate Reader with Fluorescent Read Capability
- Prepare 50 mM Sodium Borate, 1.2 M Urea, pH 8.0 by dissolving Boric Acid and Urea in deionized water and adjusting the pH with Sodium Hydroxide. Then, prepare the Assay Buffer by adding 1 M CaCl2 to a final concentration of 10 mM.
- Dilute rhLOXL2 to 10 ng/µL in Assay Buffer.
- Dilute Benzylamine to 4 mM in Assay Buffer.
- 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.
- Incubate the reactions for 30 minutes at 37 °C.
- Prepare the Substrate Mixture, 2 units/mL HRP and 40 µM AUR, in Assay Buffer.
- Load 50 µL of the incubated reactions into the wells of a plate and add 50 µL of Substrate Mixture.
- 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.
- 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 in Assay Buffer. Use this oxidized AUR curve to determine the conversion factor.
- rhLOXL2: 0.25 µg
- Benzylamine: 1 mM
- HRP: 1 unit/mL
- AUR: 20 µM
Scientific Data
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
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).
- Csiszar, H. (2001) Prog. Nucleic Acid Res. Mol. Biol. 70:1.
- Maki, J.M. and K.I. Kivirikko (2001) Biochem J. 355:381.
- Akiri, G. et al. (2003) Cancer Res. 63:1657.
- Hollosi, P. et al. (2009) Int. J. Cancer. 125:318.
- Peinado, H. et al. (2008) Cancer Res. 68:4541.
- Jourdan-Le Saux C. et al. (1999) J. Biol. Chem. 274:12939.
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.
9
Citations: Showing 1 - 9
Filter your results:
Filter by:
-
Topical application of an irreversible small molecule inhibitor of lysyl oxidases ameliorates skin scarring and fibrosis
Authors: N Chaudhari, AD Findlay, AW Stevenson, TD Clemons, Y Yao, A Joshi, S Sayar, G Wallace, S Rea, P Toshniwal, Z Deng, PE Melton, N Hortin, KS Iyer, W Jarolimek, FM Wood, MW Fear
Nature Communications, 2022-09-22;13(1):5555.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay -
LOXL2 attenuates osteoarthritis through inactivating Integrin/FAK signaling
Authors: C Zhang, M Zhu, H Wang, J Wen, Z Huang, S Chen, H Zhao, H Li
Scientific Reports, 2021-08-23;11(1):17020.
Species: Rat
Sample Types: Whole Cells
Applications: Bioassay -
The Interactome of Cancer-Related Lysyl Oxidase and Lysyl Oxidase-Like Proteins
Authors: SD Vallet, C Berthollie, R Salza, L Muller, S Ricard-Blu
Cancers, 2020-12-29;13(1):.
Species: Human
Sample Types: Recombinant Protein
Applications: Bioassay -
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, 2018-12-09;0(0):.
Applications: Bioassay -
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, 2018-11-30;17(0):38-43.
Species: Human
Sample Types: Protein
Applications: Bioassay -
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., 2017-09-05;0(0):.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay -
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, 2017-03-10;7(1):149.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
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, 2015-03-20;16(0):42.
Species: Human
Sample Types: Protein
Applications: Enzyme Assay -
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., 2011-06-11;69(1):149-63.
Species: Human, Mouse
Sample Types: Whole Cells
Applications: Enzyme Assay
FAQs
No product specific FAQs exist for this product, however you may
View all Proteins and Enzyme FAQsReviews for Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF
Average Rating: 4 (Based on 1 Review)
Have you used Recombinant Human Lysyl Oxidase Homolog 2 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by: