Recombinant Human METTL11A Protein, CF
R&D Systems | Catalog # 7159-MT
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Key Product Details
- R&D Systems E. coli-derived Recombinant Human METTL11A Protein (7159-MT)
- 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
E. coli
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
E. coli-derived human alpha-N-terminal Methyltransferase 1A/METTL11A protein
Thr2-Arg223, with an N-terminal Met and 6-His tag
Thr2-Arg223, with an N-terminal Met and 6-His tag
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Met
Predicted Molecular Mass
26 kDa
SDS-PAGE
25-26 kDa, reducing conditions
Activity
Measured by its ability to methylate the synthetic peptide Pro-Pro-Lys-Gln-Gln-Leu-Ser-Lys-Tyr.
The specific activity is >2.5 pmol/min/μg, as measured under the described conditions.
The specific activity is >2.5 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
7159-MT
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol. |
| 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.
|
Background: alpha-N-terminal Methyltransferase 1A/METTL11A
References
- Webb, K. J. et al. (2010) Biochemistry 49:5225.
- Christine, S. E. et al. (2010) Nature 466:1125.
- Chen, T. et al. (2007) Nature Cell Biol. 9:596.
- Hao, Y. et al. (2008) J. Cell. Biol. 182:827.
Long Name
alpha-N-terminal Protein Methyltransferase 1A
Alternate Names
AD-003, C9orf32, NRMT, NTM1A, NTMT1
Gene Symbol
NTMT1
UniProt
Additional alpha-N-terminal Methyltransferase 1A/METTL11A Products
Product Documents for Recombinant Human METTL11A Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human METTL11A Protein, CF
For research use only
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Protocols
View specific protocols for Recombinant Human METTL11A Protein, CF (7159-MT):
Materials
- Assay Buffer: 25 mM HEPES, 2 mM MgCl2, 1 mM EDTA, pH 8.0
- Recombinant Human alpha -N-terminal Methyltransferase 1A/METTL11A (rhMETTL11A) (Catalog # 7159-MT)
- Recombinant Human Adenosylhomocysteinase/AHCY (rhAHCY (Catalog # 6466-AH)
- Glutathione, reduced (Amresco, Catalog # 399)
- Substrate: PPKQQLSKY (SM Biochemicals, Catalog # CUS326), 1 mM stock in deionized water
- Recombinant human Adenosine Deaminase/ADA (rhADA) (Catalog # 7048-AD)
- S-Adenosylmethionine (AdoMet) (Sigma, Catalog # A7007), 10 mM stock in 50% DMSO
- ThioGlo® 3 Fluorescent Thiol Reagent (Covalent Associates Inc., Catalog # T-003)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Prepare a 10 mM stock of reduced glutathione in deionized water.
- Prepare the standard curve by diluting the 10 mM reduced glutathione (10,000 pmol/µL) to 20 pmol/µL in Assay Buffer and performing six additional ½ serial dilutions.
- Dilute rhMETTL11A to 66.67 ng/µL in Assay Buffer.
- Dilute Substrate and S-Adenosylmethionine in Assay Buffer to 133.34 μM and 666.67 uM respectively.
- Combine equal volumes of dilute Substrate and S-Adensoylmethionine for the substrate mixture. Further dilute an appropriate amount of 133.34 μM Substrate to 66.67 μM in Assay Buffer for controls.
- Dilute rhAHCY and rhADA in Assay Buffer to 50 ug/mL and 88.9 μg/mL respectively.
- Combine equal volumes of dilute rhAHCY and rhADA for the rhAHCY/rhADA mixture.
- Load the microplate as follows:
- Reactions: 15 µL of 66.67 ng/µL rhMETTL11A, 15 µL of substrate mixture, and 20 µL of rhAHCY/rhADA mixture.
- Controls: 15 µL of 66.67 ng/µL rhMETTL11A, 15 µL of 66.67 uM Substrate, and 20 µL of rhAHCY/rhADA mixture
- Standard Curve: 50 µL per well
- Seal microplate and incubate at room temperature for 30 minutes.
- Dilute ThioGlo® to 100 µM in DMSO.
- After incubation, add 50 µL of 100 µM ThioGlo® to all wells used.
- Incubate at room temperature for 5 minutes in the dark.
- Read the plate in endpoint mode at excitation and emission wavelengths of 380 nm and 445 nm, respectively.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted thiol produced* (pmol) |
| Incubation time (min) x amount of enzyme (µg) |
*Derived from the reduced glutathione standard curve using linear fitting and adjusted for Substrate Blank.
Per Well:
- rhMETTL11A: 1 μg
- rhAHCY: 0.5 µg
- Substrate: 10 µM
- S-Adenosylmethionine: 50 μM
- rhADA: 0.889 μg
- ThioGlo®: 50 µM
- Reduced Glutathione curve: 1000, 500, 250, 125, 62.5, 31.25 and 15.625 pmol