Recombinant Human AMCase/CHIA Protein, CF Summary
Product Specifications
Tyr22-Pro408, 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.
9859-GH
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with polar packs. 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 MES, pH 5.0
- Recombinant Human AMCase/CHIA (rhCHIA) (Catalog # 9859-GH)
- Substrate: 4-Methylumbelliferyl-beta -D-N, N', N''-triacetylchitotrioside (Sigma, Catalog # M5639), 5 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCHIA to 4 ng/μL in Assay Buffer.
- Dilute Substrate to 400 μM in Assay Buffer.
- Load into a plate 50 μL of 4 ng/μL rhCHIA, and start the reaction by adding 50 μL of 400 μM Substrate. Include Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 400 μM Substrate.
- Read plate at (excitation and emission) of 365 nm and 445 nm, respectively, (top read) in kinetic mode for 5 minutes.
- 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 calibration standard 4-Methylumbelliferone (Sigma, Catalog # M1381)
- rhCHIA: 0.2 μg
- Substrate: 200 μM
Reconstitution Calculator
Background: AMCase/CHIA
Human acidic mammalian chitinase (AMCase), or CHIA, degrades chitin and chitotriose, therefore participates in the defense against nematodes, fungi and other pathogens (1, 2, 3). It is one of the important proteins involved in T-helper cell type 2 (Th2)-mediated inflammation and has been implicated in asthma and allergic diseases (4). It protects lung epithelial cells against apoptosis possibly via promoting phosphorylation of AKT1 and stimulating chemokine production (5, 6). Inhibition of AMCase results in decreased airway inflammation and airway hyper-responsiveness in a mouse asthma model. Its function in the inflammatory response and in protecting cells against apoptosis is inhibited by allosamidin, suggesting that the function of this protein depends on carbohydrate binding (7).
- Boot, R.G. et al. (2001) J. Biol. Chem. 276:6770.
- Seibold, M.A. et al. (2009) J. Biol. Chem. 284:19650.
- Chou, Y.T. et al. (2006) Biochemistry 45:4444.
- Zhu, Z, et al. (2004) Science 304:1678.
- Hartl, D. et al. (2008) J. Biol. Chem. 283:33472.
- Hartl, D. et al. (2009) J. Immunol. 182:5098.
- Ober, C. and Chupp, G.L. (2009) Curr. Opin. Allergy Clin. Immunol. 9:401.
Citation for Recombinant Human AMCase/CHIA 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.
1 Citation: Showing 1 - 1
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Host gastric corpus microenvironment facilitates Ascaris suum larval hatching and infection in a murine model
Authors: Wu, Y;Adeniyi-Ipadeola, G;Adkins-Threats, M;Seasock, M;Suarez-Reyes, C;Fujiwara, R;Bottazzi, ME;Song, L;Mills, JC;Weatherhead, JE;
PLoS neglected tropical diseases
Species: Ascaris Suum
Sample Types: Egg
Applications: Bioassay
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