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- 英文名：
IDH1(R132H)
- 供应商：
倍诺博生物
- 规格：
50/100/500/1000
| 规格： | 50 | 产品价格： | 询价 |
|---|---|---|---|
| 规格： | 100 | 产品价格： | 询价 |
| 规格： | 500 | 产品价格： | 询价 |
| 规格： | 1000 | 产品价格： | 询价 |
E. coli-derived, Met1-Leu414, with a C-terminal 6-His tag
Accession #
O75874
Sequence
MSKKISGGSVVEMQGDEMTRIIWELIKEKLIFPYVELDLHSYDLGIENRDATN
DQVTKDAAEAIKKHNVGVKCATITPDEKRVEEFKLKQMWKSPNGTIRNILGG
TVFREAIICKNIPRLVSGWVKPIIIGHHAYGDQYRATDFVVPGPGKVEITYTPSD
GTQKVTYLVHNFEEGGGVAMGMYNQDKSIEDFAHSSFQMALSKGWPLYLST
KNTILKKYDGRFKDIFQEIYDKQYKSQFEAQKIWYEHRLIDDMVAQAMKSEG
GFIWACKNYDGDVQSDSVAQGYGSLGMMTSVLVCPDGKTVEAEAAHGTVT
RHYRMYQKGQETSTNPIASIFAWTRGLAHRAKLDNNKELAFFANALEEVSIE
TIEAGFMTKDLAACIKGLPNVQRSDYLNTFEFMDKLGENLKIKLAQAKL
Predicted Molecular Mass
47kDa
SDS PAGE
49kDa, reducing conditions.
Activity
Measured by the ability to convert α-ketoglutarate to 2-hydroxyglutarate. The specific activity is > 1400 pmol/min/μg, as measured under the described conditions.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Purity
>98%, by SDS-PAGE under reducing conditions and visualized by Coomassie Blue stain at 2 μg per lane.
Formulation
Supplied as a 0.2 μm filtered solution in Tris, NaCl, Glycerol, and β-mercaptoethanol.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Avoid repeated freeze-thaw cycles.
•6 months from date of receipt, -80 ◦C as supplied.
•3 months, -80 ◦C under sterile conditions after opening.
Materials
•Assay Buffer: 100 mM Tris (pH 7.5), 150 mM NaCl, 10 mM MgCl2, 1mM DTT
•Recombinant Human Isocitrate dehydrogenase 1 (mutated R132 H)/ IDH1(R132H)
•α-Ketoglutarate, 100 mM stock in deionized water
•NADPH, 50mM stock in deionized water
•96-well Clear Plate (Costar, Catalog # 2592)
•Plate Reader (Model: Tecan Infinite M200 Pro) or equivalent
Assay
1.Dilute rhIDH1(R132H) to 8 μg/mL in Assay Buffer.
2.Prepare a Substrate Mixture by Diluting NADPH and α-Ketoglutarate to 1 mM and 4mM, respectively, in Assay Buffer.
3.Load into a plate 50 μL of 8 μg/mL rhIDH1(R132H) and start the reaction by adding 50 μL of Substrate Mix. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of Substrate Mix.
4. Depletion of NADPH was monitored over time at 340 nm. Read plate at a wavelength of 340 nm (bottom read) in kinetic mode for 30 minutes.
5.Calculate specific activity:
Specific Activity (pmol/min/µg) =
*Adjusted for Substrate Blank
**Using the extinction coefficient 6220 M-1cm-1
***Using the path correction 0.32 cm
Final assay conditions
Per Well:
•rhIDH1(R132H): 0.4μg
•NADPH: 0.5 mM
•α-Ketoglutarate: 2mM
DATA
BACKGROUND
Isocitrate Dehydrogenase 1 (IDH1) catalyzes the oxidative decarboxylation of isocitrate to α ketoglutarate. The protein encoded by this gene is the NADP+-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. Mutations of Arg132 of human IDH1 result in a reduced ability of the enzyme to convert isocitrate to α-ketoglutarate, but the enzyme acquires the ability to generate 2-hydroxyglutarate (2HG) from α-ketoglutarate (1). Elevated levels of the metabolite 2HG are associated with a high risk of malignant brain tumors and other cancers such as acute myeloid leukemia.
Entrez Gene IDs:
3417 (Human); 15926 (Mouse); 24479 (Rat)
联系方式：倍诺博生物 谷小姐 13632887329/ 0755-86966133
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