Recombinant Human Chitotriosidase/CHIT1 Protein, CF Summary
Ala22-Asn466, with a C-terminal 10-His tag
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.
|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.
- 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
- Dilute rhCHIT1 to 0.4 ng/μL in Assay Buffer.
- Dilute Substrate to 200 μM in Assay Buffer.
- 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.
- Incubate at room temperature for 5 minutes. Protect from light.
- Load 90 μL of Reading Buffer per sample in plate.
- Add 10 μL of each reaction mixture into wells containing Reading Buffer.
- Immediately read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in endpoint mode.
- 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
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).
- Aguilera, B. et al. (2003) J. Biol. Chem. 278:40911.
- Boot, R.G. et al. (2001) J. Biol. Chem. 276:6770.
- Renkema, G.H. et al. (1997) Eur. J. Biochem. 244:279.
- Vellodi, A. et al. (2005) J. Inherit. Metab. Dis. 28:585.
- Sotgiu, S. et al. (2005) Eur. Neurol. 54:149.
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.
Citations: Showing 1 - 4
Filter your results:
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;11(1):19735.
Sample Types: Plasma
Applications: ELISA Standard
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;10(1):217.
Sample Types: Whole Cells
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;10(1):229.
Sample Types: Whole Tissue
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;41(4):379-84.
Sample Types: N/A
Applications: ELISA Developmet
No product specific FAQs exist for this product, however you mayView all Proteins and Enzyme FAQs
Reviews for Recombinant Human Chitotriosidase/CHIT1 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human Chitotriosidase/CHIT1 Protein, CF and earn rewards!
Have you used Recombinant Human Chitotriosidase/CHIT1 Protein, CF?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image