Recombinant Mouse IL-4I1 Protein, CF Summary
Product Specifications
Leu21-Ser630, with a C-terminal 6-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.
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.
|
Assay Procedure
- 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 range of Hydrogen Peroxide (0-500 pmol) (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.
Reconstitution Calculator
Background: IL-4I1
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).
- 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.
Citations for Recombinant Mouse IL-4I1 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.
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Citations: Showing 1 - 2
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IL-4-Induced Gene 1: A Negative Immune Checkpoint Controlling B Cell Differentiation and Activation
Authors: L Bod, L Douguet, C Auffray, R Lengagne, F Bekkat, E Rondeau, V Molinier-F, F Castellano, Y Richard, A Prévost-Bl
J. Immunol., 2017-12-29;0(0):.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay -
IL4-induced gene 1 promotes tumor growth by shaping the immune microenvironment in melanoma
Authors: L Bod, R Lengagne, L Wrobel, JP Ramspott, M Kato, MF Avril, F Castellano, V Molinier-F, A Prévost-Bl
Oncoimmunology, 2017-01-13;6(3):e1278331.
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay
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