Adenylate Kinase 1/AK1 蛋白, Rat (His)

The adenylate kinase 1 (AK1) protein plays a crucial role in cellular energy homeostasis by maintaining the balance of adenylate nucleotides by catalyzing the reversible transfer of terminal phosphate groups between ATP and AMP.Adenylate Kinase 1/AK1 Protein, Rat (His) is the recombinant rat-derived Adenylate Kinase 1/AK1 protein, expressed by E.coli , with N-His labeled tag.

腺苷酸激酶 1 (AK1) 是一种多功能酶，可催化 ATP 和 AMP 之间末端磷酸基团的可逆转移，这是细胞能量稳态的关键步骤。此外，AK1 还具有核苷二磷酸激酶活性，可使用 ATP 或 GTP 作为磷酸盐供体，促进从相应的二磷酸底物生成各种核苷三磷酸（ATP、CTP、GTP、UTP、dATP、dCTP、dGTP 和 dTTP）。这种酶的灵活性凸显了其在核苷酸生物合成中的重要作用。此外，AK1 对从二磷酸硫胺素 (ThDP) 和 ADP 合成三磷酸硫胺素 (ThTP) 的催化速率较低，表明其参与硫胺素代谢。AK1 的多方面功能强调了其在维持细胞能量水平和参与多种核苷酸和辅因子生物合成途径中的重要性。参考资料：所提供的信息基于截至 2022 年 1 月的一般知识，未引用具体参考资料。

Specific activity is 933.598 units/mg. One unit will convert ATP + AMP to 2.0 µmoles of ADP per minute at pH 7.5 at 37 °C.

Materials
Kinase-Lumi^TM Enhanced Chemiluminescent Kinase Assay Kit
Assay buffer: 40 mM Tris, 20 mM MgCl₂, 0.1 mg/mL BSA, pH 7.4
Adenylate Kinase 1/AK1 Protein, Rat (His)（HY-P74427）
Substrate: AMP, ATP
Standard: ATP

Assay Procedure
Standard Curve Preparation
1. Dilute ATP (standard) in assay buffer to the following concentrations: 0, 0.3125, 0.625, 1.25, 2.5, 5, 10, 20, 40, 50 μM.
2. Add 50 μL of each standard concentration to a well and add 50 μL of Kinase-Lumi? Enhanced Chemiluminescent Kinase Assay Reagent.
3. Incubate at room temperature in the dark for 10 minutes.
4. Using a microplate reader, measure the RLU value under luminescence.
5. Plot the measured RLU on the x-axis and the standard concentration on the y-axis to generate a standard curve and obtain the standard curve equation.

Protein Activity Assay
1. Dilute AK1 to 10 μg/mL in assay buffer.
2. Dilute ATP to 40 μM (provided in the kit) and AMP to 40 μM in assay buffer to prepare the reaction mixture.
3. Add 25 μL of 10 μg/mL AK1 to a 96-well plate, then add 25 μL of the ATP-AMP mixture to start the reaction; for the blank, add 25 μL of assay buffer and 25 μL of the ATP-AK1 mixture.
4. Incubate at 37°C in the dark for 50 minutes.
5. After incubation, add 50 μL of Kinase-Lumi^TM Enhanced Chemiluminescent Kinase Assay Reagent to each well.
6. Incubate at room temperature for 10 minutes.
7. Using a microplate reader, measure the RLU value under luminescence.
8. Calculate the specific activity:

Rat

E. coli

N-6*His

P39069 (M1-K194)

24183  [NCBI]

Full Length

MEDKLKKAKIIFVVGGPGSGKGTQCEKIVQKYGYTHLSTGDLLRAEVSSGSSRGKMLSSIMEKGELVPLETVLDMLRDAMLAKVDSSNGFLIDGYPREVKQGEEFERKIAQPTLLLYVDAGPETMTQRLLKRGETSGRVDDNEETIKKRLETYYKATEPVISFYDKRGIVRKVNAEGSVDTVFSQVCTYLDSLK

Approximately 23 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

溶液.

Supplied as a 0.22 μm filtered solution of PBS, 100 mM arginine, pH 7.1, 20% glycerol.

<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.