Recombinant Human Chitotriosidase/CHIT1 Protein, CF

R&D Systems | Catalog # 3559-GH

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Chitotriosidase/CHIT1 Protein (3559-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

NP_003456

Applications

Enzyme Activity
View all Chitotriosidase/CHIT1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Chitotriosidase/CHIT1 protein
Ala22-Asn466, with a C-terminal 10-His tag

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ala22

Predicted Molecular Mass

51 kDa

SDS-PAGE

52 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 >1,500 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

3559-GH
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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.
  • 6 months from date of receipt, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Chitotriosidase/CHIT1

Chitotriosidase encoded by the CHIT1 gene is a member of the chitinase family that is selectively expressed in activated tissue macrophages (1). It is distinct from another member, known as acidic mammalian chitinase (AMC) encoded by the CHIA gene and expressed mainly in the gastrointestinal tract and lung (2). Both CHIA and CHIT1 are secreted as 50 kDa proteins. In contrast to CHIA, CHIT1 is not stable under acidic pHs and can be processed into a C-terminally truncated 39 kDa form (2, 3). CHIT1 is the best biomarker in the monitoring of Gaucher disease among the three most commonly used markers that also include acid phosphatase and angiotensin-converting enzyme (ACE) (4). CHIT1 is also a specific marker of macrophage activation in acute ischemic stroke (5).

References

  1. Aguilera, B. et al. (2003) J. Biol. Chem. 278:40911.
  2. Boot, R.G. et al. (2001) J. Biol. Chem. 276:6770.
  3. Renkema, G.H. et al. (1997) Eur. J. Biochem. 244:279.
  4. Vellodi, A. et al. (2005) J. Inherit. Metab. Dis. 28:585.
  5. Sotgiu, S. et al. (2005) Eur. Neurol. 54:149.

Alternate Names

CHI3, CHIT1

Entrez Gene IDs

1118 (Human)

Gene Symbol

CHIT1

UniProt

NP_003456

Additional Chitotriosidase/CHIT1 Products

Product Documents for Recombinant Human 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 Human Chitotriosidase/CHIT1 Protein, CF

For research use only

Citations for Recombinant Human Chitotriosidase/CHIT1 Protein, CF

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Protocols

View specific protocols for Recombinant Human Chitotriosidase/CHIT1 Protein, CF (3559-GH):

Materials
  • Assay Buffer: 10 mM MES, pH 6.0
  • Reading Buffer: 100 mM Tris, pH 9.0
  • Recombinant Human Chitotriosidase/CHIT1 (rhCHIT1) (Catalog # 3559-GH)
  • Substrate: 4-Methylumbelliferyl beta -D-N,N’,N”-triacetylchitotriose (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 rhCHIT1 to 0.4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 200 μM in Assay Buffer.
  3. Mix equal volumes of 0.4 ng/μL rhCHIT1 and 200 μM Substrate. Include a Substrate Blank with equal volumes of Assay Buffer and Substrate.
  4. Incubate at room temperature for 5 minutes. Protect from light.
  5. Load 90 μL of Reading Buffer per sample in plate.
  6. Add 10 μL of each reaction mixture into wells containing Reading Buffer.
  7. Immediately read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in endpoint mode.
  8. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/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:
  • rhCHIT1: 0.002 μg
  • Substrate: 10 μM

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