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H+K+-ATPase Activity Assay Kit

  • Cat.No.:E-BC-K122-S

  • Detection instrument: Spectrophotometer(660 nm)

Manual MSDS

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Detection principle

ATPase can decompose ATP to produce ADP and inorganic phosphorus. The activity of ATPase can be expressed by measuring the production amount of inorganic phosphorus in unit time. The inorganic phosphorus reacts with ammonium molybdate in acidic solution to form ammonium molybdate compound, which is reduced with reducing agent to form molybdenum blue, and has absorption peak at 660 nm. Determine the concentration of molybdenum blue to calculate the amount of inorganic phosphorus.

Performance characteristics

Sample type Animal tissue,cells
Detection method Colorimetric method
Assay type Enzyme Activity
Assay time 60 min
Precision Average inter-assay CV: 9.800000000000001%Average intra-assay CV: 4.4%
Other instruments required Micropipettor, Centrifuge, Incubator, Water bath, Vortex mixer
Other reagents required Normal saline (0.9% NaCl)
Storage Reagent 4: -20℃, others: 2-8℃
Valid period 6 months

Dilution of sample

Dilute the 10% tissue homogenate to different concentrations of 2%, 1%, and 0.5%, then take 100 μL for pre-experiment according to the operation steps. The OD value of sample tube should be less than 1.0.

The recommended dilution factor for different samples is as follows (for reference only):

Sample type

Dilution factor

10% Animal tissue homogenate

5

GES-1 cells (2.52mg/mL)

1

Note: The diluent is normal saline (0.9% NaCl).

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