Recombinant Human AMCase/CHIA Protein, CF

R&D Systems | Catalog # 9859-GH

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Key Product Details

  • R&D Systems HEK293-derived Recombinant Human AMCase/CHIA Protein (9859-GH)
  • 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

HEK293

Accession Number

Q9BZP6-1

Applications

Enzyme Activity
View all AMCase/CHIA Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human AMCase/CHIA protein
Tyr22-Pro408, 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

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr22

Predicted Molecular Mass

44 kDa

SDS-PAGE

39-47 kDa, reducing conditions

Activity

Measured by its ability to cleave 4-Methylumbelliferyl-beta -D-N,N’,N”-triacetylchitotriose (4-MU-TACT).
The specific activity is >125 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

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).

References

  1. Boot, R.G. et al. (2001) J. Biol. Chem. 276:6770.
  2. Seibold, M.A. et al. (2009) J. Biol. Chem. 284:19650.
  3. Chou, Y.T. et al. (2006) Biochemistry 45:4444.
  4. Zhu, Z, et al. (2004) Science 304:1678.
  5. Hartl, D. et al. (2008) J. Biol. Chem. 283:33472.
  6. Hartl, D. et al. (2009) J. Immunol. 182:5098.
  7. Ober, C. and Chupp, G.L. (2009) Curr. Opin. Allergy Clin. Immunol. 9:401.

Long Name

Acidic Mammalian Chitinase

Alternate Names

CHIA, CHIT2, Chitinase, Acidic, Lung-Specific Protein TSA1902

Entrez Gene IDs

27159 (Human); 81600 (Mouse); 113901 (Rat)

Gene Symbol

CHIA

UniProt

Q9BZP6-1

Additional AMCase/CHIA Products

Product Documents for Recombinant Human AMCase/CHIA Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human AMCase/CHIA Protein, CF

For research use only

Citations for Recombinant Human AMCase/CHIA Protein, CF

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Protocols

View specific protocols for Recombinant Human AMCase/CHIA Protein, CF (9859-GH):

Materials
  • 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
  1. Dilute rhCHIA to 4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 400 μM in Assay Buffer.
  3. 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.
  4. Read plate at (excitation and emission) of 365 nm and 445 nm, respectively, (top read) in kinetic mode for 5 minutes.
  5. 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)

Per Well:
  • rhCHIA: 0.2 μg
  • Substrate: 200 μM

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