ACOT13 Protein, Human (HEK293, His)

ACOT13 Protein, Human (HEK293, His) is human recombinant Acyl-coenzyme A thioesterase 13 (ACOT13) with a N-terminal His-tag. ACOT13 Protein, Human (HEK293, His) contains 160 amino acids (1-140 a.a.).

Acyl-coenzyme A thioesterase 13 (ACOT13), also known as thioesterase superfamily member 2 (Them2), belongs to the mammalian acyl-CoA thioesterase family. ACOT13 is a homotetrameric hotdog fold thioesterase, and is enriched in oxidative tissues, associated with mitochondria, and relatively specific for long chain fatty acyl-CoA substrates. ACOT13 plays a key role in hepatic lipid and glucose metabolism. ACOT13 is essential for the cell sustained proliferation and is co-localized with microtubules^[1][2].

Measured by its ability to Hydrolysis 500μg/mL Myristoyl coenzyme A lithium and generate product of TNB. The specific activity is 6.66 pmol/min/µg that incubate at 25 ℃ in kinetic mode for 60 minutes.

Materials
Assay Buffer: 50 mM KCl, 10 mM Hepes and 0.3 mM DTNB , pH 7.5
Substrate: Myristoyl coenzyme A lithium (HY-126833A)
ACOT13 Protein, Human (HEK293, His) (HY-P7452)
Standard: TNB

Procedure
1. Dilute Human ACOT13 to 100 μg/mL in experimental buffer.
2. Dilute the substrate to 1 mg/mL in the experimental buffer.
3. Experimental group: 50 μL Human ACOT13 protein + 50 μL substrate.
Blank group: 50 μL experimental buffer + 50 μL substrate.
4. Determination of OD value at 412 nm for 1 hour under kinetic mode.
5. Preparation of standard curve: Dilute TNB with experimental buffer to a final concentration of 0, 0.0001, 0.001, 0.01, 0.1, 1 mM, 100 μL per well was added into 96-well plate, and OD₄₁₂ was measured in endpoint mode.
6. Calculate specific activity:

Human

HEK293

C-6*His

Q9NPJ3-1 (T2-N140)

55856  [NCBI]

Full Length of Mature Protein

TSMTQSLREVIKAMTKARNFERVLGKITLVSAAPGKVICEMKVEEEHTNAIGTLHGGLTATLVDNISTMALLCTERGAPGVSVDMNITYMSPAKLGEDIVITAHVLKQGKTLAFTSVDLTNKATGKLIAQGRHTKHLGN

Approximately 15.9 kDa

≥ 95%, as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 8% Sucrose, 100 mM NaCl, 0.05% Tween 80, pH 7.0.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

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.

• [1]. Wei J, et al. Thioesterase superfamily member 2 (Them2)/acyl-CoA thioesterase 13 (Acot13): a homotetrameric hotdog fold thioesterase with selectivity for long-chain fatty acyl-CoAs. Biochem J. 2009 Jun 26;421(2):311-22.  [Content Brief]

  [2]. Cheng Z, et al. Human thioesterase superfamily member 2 (hTHEM2) is co-localized with beta-tubulin onto the microtubule. Biochem Biophys Res Commun. 2006 Dec 1;350(4):850-3.  [Content Brief]