Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 6 hr
- Sample type: Cell culture extracts, Tissue
Overview
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Product name
Complex IV Rodent Enzyme Activity Microplate Assay Kit
See all Complex IV kits -
Detection method
Colorimetric -
Sample type
Cell culture extracts, Tissue -
Assay type
Enzyme activity (quantitative) -
Assay time
6h 00m -
Species reactivity
Reacts with: Mouse, Rat -
Product overview
Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911) is used to determine the activity of cytochrome c oxidase in a mouse sample with speed and simplicity. The COX enzyme is immunocaptured within the wells of the microplate and activity is determined colorimetrically by following the oxidation of reduced cytochrome c by the absorbance change at 550 nm. Included in this kit for performance of the activity assay are buffer, detergent, substrate, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.
Complex IV assay protocol summary:
- add samples to wells to capture enzyme and incubate for 3 hrs
- wash wells
- add reaction mix
- analyze with microplate reader in kinetic mode for 120 minBuffer, detergent, and microplate should be stored at 4°C, Reagent C (reduced cytochrome c) should be stored at -80°C.
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Notes
Range of complex IV / cytochrome c oxidase assay kits
Biochemical assay - ab239711
Immunocapture with biochemical assay (plate-based)*** - ab109911 (rodent) (this kit) and ab109909 (human)
*** Most popular assay formatImmunocapture with biochemical assay (dipstick) - ab109878 (rodent) and ab109876 (human)
Immunocapture with biochemical assay and ELISA - ab109910 (human)
ELISA - ab179880 (human)
Other related productsReview the mitochondrial assay guide, or the full metabolism assay guide 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.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 96 tests 96-well microplate 1 unit Tube 1 (Buffer) 1 x 10ml Detergent 1 x 1ml Reagent C (Reduced Cytochrome c) 1 x 1ml -
Research areas
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Alternative names
- cco
- COX
- Cytochrome c oxidase
- Cytochrome oxidase
Images
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Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)Figure 1. To determine the activity in the sample, calculate the slope by using microplate software or by manual calculations using one of the two methods shown. Compare the sample rate with the rate of the control (normal) sample and with the rate of the null (background) to get the relative Complex IV activity. (A)The rate is determined by calculating the gradient of the initial slope over the linear region. (B) The rate is determined by calculating the slope between two points within the linear region.
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Functional Studies - Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911)Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states. Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, substrate is added, and enzyme activity is analyzed by measuring the change in absorbance of either the substrate or the product of the reaction (depending upon which enzyme is being analyzed). 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. -
Mitochondrial ROS dependent functional deficiency and structural impairment in cardiac mitochondria after sepsis. Image courtesy of Yao X et al.PLoS One. 2015; 10(10): e0139416. doi: 10.1371/journal.pone.0139416.Mitchondiral fractions from the heart tissue of Rats infected by S. pneumoniae, or given PBS sham control, were subjected to measurements of complex I-V activities. Complex I was measured with ab109721 (top left), Complex II + III were measured using ab109905 (top right), Complex IV was measured using ab109911 (bottom left) and Complex V was measured using ab109714 (bottom right).
Freshly isolated mitochondrial pellets were resuspened in PBS supplemented with 10% detergent provided in the kits. Protein concentrations of these mitochondrial lysates were estimated and 25 μg (for complex I, IV and V) or 100 μg (for complex II+III) mitochondrial protein was used per reaction. Enzyme activities were measured spectrophotometricly in triplicate and expressed as changes of absorbance per minute per mg protein.
