Recombinant Mouse Chitotriosidase/CHIT1 Protein, CF

R&D Systems | Catalog # 5325-GH

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse Chitotriosidase/CHIT1 Protein (5325-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

NS0

Accession Number

Q9D7Q1

Applications

Enzyme Activity
View all Chitotriosidase/CHIT1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Chitotriosidase/CHIT1 protein
Ala22-Ser464 (major) & Lys23-Ser464 (minor), both with a C-terminal 6-His tag

Purity

>90%, 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

Ala22 & Lys23

Predicted Molecular Mass

50 kDa

SDS-PAGE

54-57 kDa doublet, reducing conditions

Activity

Measured by its ability to cleave 4-Methylumbelliferyl-beta -D-N,N’,N”-triacetylchitotriose (4-MU-TACT).
The specific activity is >400 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5325-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: Chitotriosidase/CHIT1

Chitotriosidase, encoded by the mouse CHIT1 gene, is a typical member of the chitinase family (1). It is distinct from another member known as acidic mammalian chitinase (AMC), encoded by the CHIA gene, in several aspects. AMC/CHIA is expressed mainly in alveolar macrophages and in both the mouse and human gastrointestinal tract (2). CHIT1, however, is expressed exclusively by phagocytes in humans and in the gastrointestinal tract, tongue, fore-stomach, and Paneth cells of the small intestine in mice (2). Both CHIA and CHIT1 are secreted as 50 kDa proteins. In contrast to CHIA, CHIT1 is not stable under acidic pH and can be processed into a C-terminally truncated 39 kDa form (2‑4). Human CHIT1 is the best of three biomarkers in the monitoring of Gaucher disease (5). The other two commonly used markers include acid phosphatase and angiotensin-converting enzyme (ACE) (5). Human CHIT1 is also a specific marker of macrophage activation in acute ischemic stroke (6).

References

  1. Zheng, T. et al. (2005) Gene 357:37.
  2. Boot, R.G. et al. (2006) J. Histochem. Cytochem. 53:1283.
  3. Boot, R.G. et al. (2001) J. Biol. Chem. 276:6770.
  4. Renkema, G.H. et al. (1997) Eur. J. Biochem. 244:279.
  5. Vellodi, A. et al. (2005) J. Inherit. Metab. Dis. 28:585.
  6. Sotgiu, S. et al. (2005) Eur. Neurol. 54:149.

Alternate Names

CHI3, CHIT1

Entrez Gene IDs

1118 (Human)

Gene Symbol

CHIT1

UniProt

Q9D7Q1

Additional Chitotriosidase/CHIT1 Products

Product Documents for Recombinant Mouse Chitotriosidase/CHIT1 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Chitotriosidase/CHIT1 Protein, CF

For research use only

Citations for Recombinant Mouse Chitotriosidase/CHIT1 Protein, CF

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Protocols

View specific protocols for Recombinant Mouse Chitotriosidase/CHIT1 Protein, CF (5325-GH):

Materials
  • Assay Buffer: 10 mM MES, 0.05% (w/v) Brij-35, pH 6.0
  • Reading Buffer: 100 mM Tris, pH 9.0
  • Recombinant Mouse Chitotriosidase/CHIT1 (rmCHIT1) (Catalog # 5325-GH)
  • Substrate: 4-Methylumbelliferyl beta -D-N’,N”-triacetylchitotrioside (4-MU-TACT) (Sigma, Catalog # M-5639), 5 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmCHIT1 to 2 µg/mL in Assay Buffer.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Mix 20 µL of 2 µg/mL of rmCHIT1 and 20 µL of 200 µM of Substrate. Include a Substrate Blank containing 20 µL Assay Buffer and 20 µL Substrate.
  4. Incubate at room temperature for 10 minutes. Protect from light.
  5. Load 90 µL of Reading Buffer per sample into a plate.
  6. Add 10 µL from each reaction mixture into wells containing Reading Buffer.
  7. Read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in endpoint mode.
  8. Calculate specific activity:

     Specific Activity (pmoles/min/µg) =

 Adjusted Fluorescence* (RFU) x Conversion Factor** (pmole/RFU)
Incubation time (min) x amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Methylumbelliferone (4-MU) (Sigma, Catalog # M1381).

Per Well:
  • rmCHIT1: 0.01 µg
  • Substrate: 10 µM

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