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Complex IV Human Enzyme Activity Dipstick Assay Kit is an immunoassay kit for the detection of Complex IV Human Enzyme Activity in Cow, Human in Cell culture extracts, Tissue samples.

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Sample types

Cell culture extracts, Tissue

Assay type

Sandwich (quantitative)

Reactive species

Cow, Human

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Reactivity data

Application
FuncS
Reactivity
Reacts
Dilution info
-
Notes

-

Images

Target data

Function

Component of the cytochrome c oxidase, the last enzyme in the mitochondrial electron transport chain which drives oxidative phosphorylation. The respiratory chain contains 3 multisubunit complexes succinate dehydrogenase (complex II, CII), ubiquinol-cytochrome c oxidoreductase (cytochrome b-c1 complex, complex III, CIII) and cytochrome c oxidase (complex IV, CIV), that cooperate to transfer electrons derived from NADH and succinate to molecular oxygen, creating an electrochemical gradient over the inner membrane that drives transmembrane transport and the ATP synthase. Cytochrome c oxidase is the component of the respiratory chain that catalyzes the reduction of oxygen to water. Electrons originating from reduced cytochrome c in the intermembrane space (IMS) are transferred via the dinuclear copper A center (CU(A)) of subunit 2 and heme A of subunbit 1 to the active site in subunit 1, a binuclear center (BNC) formed by heme A3 and copper B (CU(B)). The BNC reduces molecular oxygen to 2 water molecules using 4 electrons from cytochrome c in the IMS and 4 protons from the mitochondrial matrix.

What's included?

30 Test
Components
96-well microplate
2 x 1 Unit
Buffer B (10X Blocking solution)
2 x 1 mL
Dipsticks
1 x 30 Unit
Extraction Buffer (ab260490)
1 x 15 mL
Tube 1 (DAB 100X stock)
1 x 100 µL
Tube 2 (reduced cytochrome c 20X stock)
1 x 500 µL
Tube 3 (Buffer for activity assay)
1 x 10 mL
Wash buffer
1 x 1.5 mL

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Complex IV Human Enzyme Activity Dipstick Assay Kit is an immunoassay kit for the detection of Complex IV Human Enzyme Activity in Cow, Human in Cell culture extracts, Tissue samples.

Key facts

Sample types

Cell culture extracts, Tissue

Assay type

Sandwich (quantitative)

Reactive species

Cow, Human

Assay Platform

Reagents

Storage

Shipped at conditions

Dry Ice

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

Contains 30 or 90 dipsticks and necessary components to quantify the activity of the cytochrome c oxidase enzyme complex from human and bovine samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.


The isolation of mitochondria is not necessary for the performance of this assay. In this kit the specificity of anti-COX monoclonal antibodies (mAbs) is combined with traditional methods for determining COX enzyme activity by histochemical methods and in-gel activity assays. First, the COX enzyme complex is immnocaptured (i.e immunoprecipated in active form) on the dipstick. Second, the dipstick is immersed in COX activity buffer containing reduced cytochrome c and di-amino benzidinetetrachloride (DAB), which serves as the reporter of COX activity. Immunocaptured COX oxidizes cytochrome c, which then oxidizes DAB to form a red-colored precipitate at the COX antibody line on the dipstick. In addition to being quick, the reaction is cyanide-sensitive. The signal intensity of this precipitate corresponds to the level of COX activity in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.

Store dipsticks at room temperature in their provided container and out of direct sunlight. High humidity conditions should be avoided.

Store Buffer A, B, and C at 4°C or at -20°C for long term storage.

Store Tubes 1 and 2 at -80°C; they can also be aliquoted upon receipt to prevent freeze/thaw cycles.

Tube 3 can be stored at room temperature.

Range of complex IV / cytochrome c oxidase assay kits

Biochemical assay - ab239711

Immunocapture with biochemical assay (plate-based) - ab109911 (rodent) and ab109909 (human)

Immunocapture with biochemical assay (dipstick) - ab109878 (rodent) and ab109876 (human) (this kit)

Immunocapture with biochemical assay and ELISA - ab109910 (human)

ELISA - ab179880 (human)

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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