AMCase/CHIA Protein, Human (HEK293, His)
Based on 1 Customer Validation
AMCase/CHIA protein is an enzyme that degrades chitin and chitotriose and is involved in defense against nematodes, fungi, and pathogens.It plays a role in Th2 immune responses, in response to IL-13, and in the induction of inflammation.AMCase/CHIA Protein, Human (HEK293, His) is the recombinant human-derived AMCase/CHIA protein, expressed by HEK293 , with C-His labeled tag.
- Species: Human
- Source: HEK293
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Storage:Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
AMCase/CHIA protein is an enzyme that degrades chitin and chitotriose and is involved in defense against nematodes, fungi, and pathogens.It plays a role in Th2 immune responses, in response to IL-13, and in the induction of inflammation.AMCase/CHIA Protein, Human (HEK293, His) is the recombinant human-derived AMCase/CHIA protein, expressed by HEK293 , with C-His labeled tag.
Background
The AMCase/CHIA protein functions as an enzyme that degrades chitin and chitotriose, suggesting its involvement in defense mechanisms against nematodes, fungi, and other pathogens. Additionally, AMCase/CHIA plays a role in T-helper cell type 2 (Th2) immune response and contributes to the response to IL-13, as well as inflammation induced by IL-13. It stimulates the production of chemokines by pulmonary epithelial cells, showcasing its influence on immune and inflammatory responses. Moreover, AMCase/CHIA is implicated in the protection of lung epithelial cells against apoptosis, promoting the phosphorylation of AKT1. Notably, the protein's function in the inflammatory response and cell protection against apoptosis is inhibited by allosamidin, suggesting a dependency on carbohydrate binding for its proper functioning. The diverse roles of AMCase/CHIA underscore its significance in immune defense, inflammation, and cellular homeostasis.
Verified Bioactivity
Measured by its ability to cleave 400 μM 4-Methylumbelliferyl-beta -D-N,N’,N”-triacetylchitotriose (4-MU-TACT). The specific activity is 371.38 pmol/min/µg.
Assay Procedure
Materials
Assay buffer: 0.1 M MES, pH 5.0
AMCase/CHIA Protein, Human (HEK293, His) (HY-P77869)
Substrate: 4-methylumbelliferyl-β-D-N,N',N'-triacetylchitotrioside, prepared as a 5 mM stock solution in DMSO
Procedure
1. Dilute Human CHIA to 4 ng/μL in assay buffer.
2. Dilute the substrate to 400 μM in assay buffer.
3. Add 50 μL of 4 ng/μL Human CHIA to a black 96-well plate, followed by 50 μL of 400 μM substrate to initiate the reaction. Include a substrate blank with 50 μL of assay buffer and 50 μL of 400 μM substrate. Perform three replicates for both the experimental and control groups.
4. Read the fluorescence in kinetic mode for 5 minutes at excitation and emission wavelengths of 365 nm and 445 nm, respectively, to obtain RFU values.
5. Standard Preparation: Prepare standard dilutions in assay buffer with concentrations of 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, 3.13 μM, 1.56 μM. Perform three replicates for each concentration.
6. Calculate enzyme 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
Per Well:
Rat TACE/ADAM17: 0.2 μg
Substrate: 200 μM
Technical Parameters
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Species Human
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Source HEK293
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Tag C-8*His
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Accession
Q9BZP6 (Y22-A476)
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Molecular Construction
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N-term
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CHIA (Y22-A476)
Accession # Q9BZP6 -
8*His
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C-term
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Protein Length
Full Length of Mature Protein
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Synonyms
CHIA; TSA1902; Chitinase Acidic; CHIT2; Acidic Mammalian Chitinase; Lung-Specific Protein TSA1902; AMCase; Chitinase, Acidic
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AA Sequence
YQLTCYFTNWAQYRPGLGRFMPDNIDPCLCTHLIYAFAGRQNNEITTIEWNDVTLYQAFNGLKNKNSQLKTLLAIGGWNFGTAPFTAMVSTPENRQTFITSVIKFLRQYEFDGLDFDWEYPGSRGSPPQDKHLFTVLVQEMREAFEQEAKQINKPRLMVTAAVAAGISNIQSGYEIPQLSQYLDYIHVMTYDLHGSWEGYTGENSPLYKYPTDTGSNAYLNVDYVMNYWKDNGAPAEKLIVGFPTYGHNFILSNPSNTGIGAPTSGAGPAGPYAKESGIWAYYEICTFLKNGATQGWDAPQEVPYAYQGNVWVGYDNIKSFDIKAQWLKHNKFGGAMVWAIDLDDFTGTFCNQGKFPLISTLKKALGLQSASCTAPAQPIEPITAAPSGSGNGSGSSSSGGSSGGSGFCAVRANGLYPVANNRNAFWHCVNGVTYQQNCQAGLVFDTSCDCCNWA
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Predicted Molecular Mass
51.2 kDa
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Molecular Weight
Approximately 55-60 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.
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Purity
≥ 95%, as determined by Bis-Tris PAGE.
Product Properties
Lyophilized powder
Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
<1 EU/μg, determined by LAL method.
It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.
Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
Room temperature in continental US; may vary elsewhere.
Documentation
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Data Sheet (238 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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