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Recombinant Human Chitotriosidase/CHIT1 Protein, CF

Catalog #: 3559-GH
4 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
3559-GH-010
R&D Systems Recombinant Proteins and Enzymes
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Product Details
Citations (4)
FAQs
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Chitotriosidase/CHIT1 Protein, CF Summary

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived human Chitotriosidase/CHIT1 protein
Ala22-Asn466, with a C-terminal 10-His tag
Accession #
NP_003456
N-terminal Sequence
Analysis
Ala22
Predicted Molecular Mass
51 kDa
SDS-PAGE
52 kDa, reducing conditions

Product Datasheets

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3559-GH

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
1118 (Human)
Alternate Names
CHI3; CHIT1; CHITCHI3; chitinase 1 (chitotriosidase); chitinase-1; Chitotriosidase; chitotriosidase-1; EC 3.2.1.14; FLJ00314; MGC125322; plasma methylumbelliferyl tetra-N-acetylchitotetraoside hydrolase

Citations for Recombinant Human Chitotriosidase/CHIT1 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.

4 Citations: Showing 1 - 4
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  1. Dysregulated non-coding telomerase RNA component and associated exonuclease XRN1 in leucocytes from women developing preeclampsia-possible link to enhanced senescence
    Authors: T Lekva, MCP Roland, ME Estensen, ER Norwitz, T Tilburgs, T Henriksen, J Bollerslev, KR Normann, P Magnus, OK Olstad, P Aukrust, T Ueland
    Scientific Reports, 2021-10-05;11(1):19735.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Standard
  2. Neuroinflammation-induced lymphangiogenesis near the cribriform plate contributes to drainage of CNS-derived antigens and immune cells
    Authors: M Hsu, A Rayasam, JA Kijak, YH Choi, JS Harding, SA Marcus, WJ Karpus, M Sandor, Z Fabry
    Nat Commun, 2019-01-16;10(1):229.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  3. Chi3l3 induces oligodendrogenesis in an experimental model of autoimmune neuroinflammation
    Authors: SC Starossom, J Campo Garc, T Woelfle, S Romero-Sua, M Olah, F Watanabe, L Cao, A Yeste, JJ Tukker, FJ Quintana, J Imitola, F Witzel, D Schmitz, M Morkel, F Paul, C Infante-Du, SJ Khoury
    Nat Commun, 2019-01-15;10(1):217.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. Macrophage chitinase 1 stratifies chronic obstructive lung disease.
    Authors: Agapov E, Battaile JT, Tidwell R, Hachem R, Patterson GA, Pierce RA, Atkinson JJ, Holtzman MJ
    Am. J. Respir. Cell Mol. Biol., 2009-06-02;41(4):379-84.
    Applications: ELISA Developmet

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