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Complex I Enzyme Activity Assay Kit (ab109721) is a kit designed for the analysis of mitochondrial OXPHOS Complex I enzyme activity from human, rat, mouse and bovine cell and tissue extracts.

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue

Assay type

Enzyme activity

Reactive species

Mouse, Rat, Cow, Human

Assay time

3h 30m

Images

What's included?

96 Test
Components
100X Dye (lyophilized)
1 x 1 Unit
10X Blocking Solution
1 x 10 mL
20X NADH (lyophilized)
1 x 1 Unit
20X Wash Buffer
1 x 25 mL
96-well microplate (12 strips)
1 x 1 Unit
Detergent
1 x 1 mL

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Complex I Enzyme Activity Assay Kit (ab109721) is a kit designed for the analysis of mitochondrial OXPHOS Complex I enzyme activity from human, rat, mouse and bovine cell and tissue extracts.

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue

Assay type

Enzyme activity

Reactive species

Mouse, Rat, Cow, Human

Assay time

3h 30m

Assay Platform

Microplate reader

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Complex I Enzyme Activity Assay Kit (ab109721) is a kit designed for the analysis of mitochondrial OXPHOS Complex I enzyme activity from human, rat, mouse and bovine cell and tissue extracts. Capture antibodies specific for Complex I are pre-coated in the microplate wells. Samples are added to the microplate wells which have been precoated with a specific capture antibody. After the target has been immobilized in the well, Complex I activity is determined by following the oxidation of NADH to NAD+ and the simultaneous reduction of a dye which leads to increased absorbance at OD=450 nm. By analyzing the enzyme's activity in an isolated context, outside of the cell and free from any other variables, an accurate measurement of the enzyme's functional state can be understood. Complex I activity is controlled by enzyme amount and by post-translational phosphorylation at key specific regulatory residues. Cellular metabolism governs these two factors. Ultimately, the cell type and growth conditions will affect Complex I activity measurements. Note: This activity assay measures the diaphorase-type activity of Complex I. This activity is not dependant on the presence of ubiquinone and therefore inhibitors, such as rotenone, which bind at or near the ubiquinone binding site do not inhibit this assay. However, both the activity assay and the quantity assay described here are affected by enzyme assembly deficiencies. Review our to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader. **Chinese protocol available.** See protocols section below.

ab109721 is shipped at 4°C. 20X NADH and 100X Dye are shipped lyophilized. Rehydrate 20X NADH by adding 1.1 mL H2O. Rehydrate 100X Dye by adding 0.25 mL H2O, then vortex each thoroughly until dissolved. After hydration unused amounts of these two materials should be stored at -80°C. When planning multiple experiments, aliquot these reagents to prevent freeze thaw cycles. Store all other components at 4°C **Related products** Review the , or the full to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

Product promise

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Full details and terms and conditions can be found here:
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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 PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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