ATPase Assay Kit (Colorimetric)

• FuncS

Supplier Data

Functional Studies - ATPase Assay Kit (Colorimetric) (AB234055)

Specific ATPase Activity were calculated in lysates prepared from Rat Heart (35 µg), Rat Kidney (15 µg), and Hela Cell Lysate (5.4 µg)

• FuncS

Supplier Data

Functional Studies - ATPase Assay Kit (Colorimetric) (AB234055)

Phosphate Standard Curve

• Biochemical assay

PubMed

Biochemical assay - ATPase Assay Kit (Colorimetric) (AB234055)

Khilazheva et al used Lactate assay kit ab65330 and ATPase assay kit ab234055 to investigate how an inducible Slc4a11 KO mouse leads to corneal edema by disruption of the pump and barrier functions of the corneal endothelium (CE).

Changes in the pump function in the inducible Slc4a11 KO. Stomal lactate content; relative values versus control; mean ± SEM, n = 3, * : p < 0.05. Na+-K+ ATPase activity at 14 days of Tm treatment; mean ± SEM, n = 3 (Ctrl) and n = 5 (Tm), * : p < 0.05.

Stromal Lactate

Corneas were obtained, and the epithelium and endothelium were removed. Individual stromas were placed in pre-weighed Eppendorf tubes, pulverized in liquid nitrogen, and homogenized in 30 μL of PBS using a plastic disposable pestle. The sample was centrifuged at 15,000x g for 15 min at 4 °C. The supernatant was recovered. The remaining pellet was dried at 60 °C in a vacuum centrifuge for two hours, then weighed (dry weight). Lactate was measured in the supernatant, n 3 for Ctrl and Tm, using a fluorescent kit (Abcam #ab65330, Cambridge, UK) according to the manufacturer's instructions.

Na+-K+ ATPase Activity

Two corneal endothelial-Descemet membrane (CEDM) peelings from the same mouse were pooled and homogenized in 30 μL assay buffer, provided in the ATPase Assay kit (Abcam #ab234055), using a plastic disposable pestle. After sonication, the sample was centrifugated at 10,000x g for 10 min at 4 °C. The supernatant was recovered, and the phosphates in the sample were depleted by incubation in 40 μL of PiBind resin (Innova Biosciences #501-0015, Montluçon, France) for 15 min at room temperature in a rotary device. After centrifugation at 1000x g for two minutes, the sample was recovered, and ATPase activity was measured in 5 μL of sample in the presence or absence of 1 mM ouabain. Na+-K+ ATPase activity (n 3 for Ctrl and Tm) was obtained by subtracting the activity in presence of ouabain from the total activity. Protein was measured using the BCA method.

• Other

Supplier Data

Other - ATPase Assay Kit (Colorimetric) (AB234055)

Representative image of ATPase Assay Kit (Colorimetric) ab234055

Components shown from left to right :

- ATP IV (2 vials)

- Phosphate Standard

- ATPase Positive Control

- ATPase Developer

- ATPase Assay Buffer

Note : The vial labels shown in this image use generic names for illustrative purposes only and may not exactly match the specific component names included in the kit.

Note : Colors of solutions in image may not precisely match the shade of colors in the actual kit.