Aldolase B 蛋白, Human (GST, solution)

Aldolase B protein catalyzes fructose 1,6-bisphosphate to trisaccharide phosphate during glycolysis and performs reverse aldol addition during gluconeogenesis. Aldolase B Protein, Human (GST, solution) is the recombinant human-derived Aldolase B protein, expressed by E. coli , with N-GST labeled tag.

醛缩酶 B 蛋白通过催化果糖 1,6-二磷酸羟醛裂解为磷酸二羟丙酮和 D-甘油醛 3-磷酸，以及糖异生中的反向立体特异性羟醛加成反应，在糖酵解中发挥关键作用。此外，在果糖分解过程中，它将膳食果糖通过果糖激酶磷酸化而产生的 1-磷酸果糖代谢为磷酸二羟丙酮和 D-甘油醛。除了其酶促功能外，醛缩酶 B 还充当接头，并通过稳定与 G6PD 和 TP53 的三元复合物来发挥肿瘤抑制因子的作用，从而抑制 G6PD 活性。这种调节机制有助于维持对氧化戊糖磷酸代谢的控制，有助于细胞代谢途径的整体平衡。

The specific activity is 1.3853 U/mg .One unit will convert 1.0 µmol of fructose 1,6-diphosphate to dihydroxyacetone phosphate and glyceraldehydes 3- phosphate per minute at pH 7.5 at 37 °C.

Materials
Substrate: Accurately weigh 16.24 mg of substrate powder and dissolve in 1 mL of deionized water to achieve a concentration of 40 mM.
Aldolase B Protein, Human (GST, solution) (HY-P74419)
NADH: Accurately weigh 5.672 mg of solid NADH powder and dissolve in 1 mL of deionized water to achieve a concentration of 8 mM.

Procedure
1. Serially dilute NADH to the following concentrations: 0, 0.0150625, 0.03125, 0.0625, 0.125, 0.25, 0.5, 1, 2 mM. Add 200 μL per well to a 96-well plate with three replicates.
2. Measure the absorbance at 340 nm using a microplate reader. Plot a standard curve with the amount of NADH as the abscissa and OD value as the ordinate.
3. Dilute the substrate to 40 mM and set aside.
4. Mix 50 μL of 40 mM substrate with 50 μL of 1 mM NADH, add to the 96-well plate in advance, and set three replicates.
5. Dilute Human Aldolase B to 0.01 mg/mL. Initiate the reaction by adding 100 μL per well.
6. Zero the microplate reader using the OD value of the blank group, and read the absorbance at 340 nm for 10 min in kinetic mode.
7. Define one enzyme activity unit (U) as the amount that consumes 1 μmol of NADH per minute. Calculate the specific enzyme activity:

Human

E. coli

N-GST

P05062 (A2-Y364)

229  [NCBI]

Full Length of Mature Protein

Approximately 60 KDa

• 
  Greater than 90% as determined by reducing SDS-PAGE.

溶液.

Supplied as a 0.22 μm filtered solution of PBS, pH 7.5.

<1 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.