Interleukin four-induced gene 1 (IL-4I1) was first identified in mouse B cells as part of the response to IL-4, a key regulator of the immune response (1). Later, the enzyme was shown to be a member of the flavin-containing amino acid oxidase family and closely related to L-amino acid oxidases (2, 3). Enzymological characterization reveals that IL-4I1 has L-amino acid oxidase activity with preference toward aromatic amino acids (4). Its limited expression in immune tissues, dendritic cells, and macrophages indicates that it may play an important function in the immune system (5, 6).
Recombinant Mouse IL-4I1 Protein, CF
R&D Systems | Catalog # 5576-AO
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Key Product Details
- R&D Systems CHO-derived Recombinant Mouse IL-4I1 Protein (5576-AO)
- 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
CHO
Accession Number
Applications
Bioactivity
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Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived mouse IL-4I1 protein
Leu21-Ser630, with a C-terminal 6-His tag
Leu21-Ser630, with a C-terminal 6-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
Leu21
Predicted Molecular Mass
69 kDa
SDS-PAGE
86 kDa, reducing conditions
Activity
Measured by its ability to produce hydrogen peroxide during the oxidation of phenylalanine.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
The specific activity is >100 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
5576-AO
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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.
|
Background: IL-4I1
References
- Chu, C.C. and W.E. Paul (1997) Proc. Natl. Acad. Sci. USA 94:2507.
- Raibekas, A.A. and V. Massey (1998) Biochem. Biophys. Res. Comm. 248:476.
- Chavan, S.S. et al. (2002) Biochim. Biophys. Acta 1576:70.
- Mason, J.M. et al. (2004) J. Immunol. 173:4561.
- Copie-Berman, C. et al. (2003) Blood 101:2756.
- Bolland, M.-L. et al. (2007) Blood 110:220.
Long Name
Interleukin 4-induced Protein-1
Alternate Names
FIG1, IL4I1
Gene Symbol
IL4I1
UniProt
Additional IL-4I1 Products
Product Documents for Recombinant Mouse IL-4I1 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.
Product Specific Notices for Recombinant Mouse IL-4I1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Mouse IL-4I1 Protein, CF
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Protocols
View specific protocols for Recombinant Mouse IL-4I1 Protein, CF (5576-AO):
Materials
- Assay Buffer: 0.1 M Sodium Phosphate, pH 8.0
- Recombinant Mouse IL-4I1 (rmIL-4I1) (Catalog # 5576-AO)
- Coupling Agent: 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: Phenylalanine (Sigma, Catalog # P2126), 100 mM stock in diH2O
- 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
- Dilute rmIL-4I1 to 2 ng/µL in Assay Buffer.
- Prepare the Substrate mixture including 2 mM Phenylalanine, 2 units/mL HRP and 100 µM AUR in Assay Buffer.
- Load in a black well plate 50 µL of 2 ng/µL of rmIL-4I1, and start the reaction by adding 50 µL of the Substrate mixture (step 2). Include a Substrate Blank containing 50 µL of the Assay Buffer and 50 µL of the Substrate mixture.
- Read at excitation and emission wavelengths of 544 nm and 590 nm (top read), respectively in kinetic mode for 5 minutes. Note: A wavelength cutoff of 570 nm should be manually specified.
- 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 a fluorescent standard prepared
by incubating 50 µM AUR, 1 unit/mL HRP, 1 mM Phenylalanine, and a curve of
Hydrogen Peroxide (Sigma, Catalog # H1009) in Assay Buffer at room temperature.
Use the resulting amounts of oxidized AUR to determine the conversion factor.
- rmIL-4I1: 0.1 µg
- Phenylalanine: 1 mM
- HRP: 1 unit/mL
- AUR: 50 µM
Caution: The rmIL-4I1 enzymatic reaction may significantly be interfered with organic buffer such as MES.
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