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Products:Signal Transduction >> Metabolism >> Mitochondrial
MSCatalog No. MS432-30
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Read our guarantee »Complex IV Rodent Enzyme Activity Dipstick Assay Kit
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Cell culture extracts, Tissue
Quantitative
Reacts with
Mouse, Rat
ab109878(MS432) contains 30 or 90 dipsticks and necessary components to quantify the activity of the cytochrome c oxidase enzyme complex from mouse and rat samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.
Using an antibody immobilized on the dipstick that specifically recognizes COX, the enzyme complex is immno-captured (i.e immuno-precipitated in active form) and enzyme activity is determined directly on the dipstick. Using a traditional method for determining COX enzyme activity by histo-chemical methods and in-gel activity assays, Di-amino benzidinetetrachloride (DAB) serves as the reporter of COX activity. The greater the signal of precipitated DAB, the greater the amount of COX enzyme complex. In addition, the reaction is cyanide sensitive. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.
Functional Studiesmore details
Please see Notes section
| Components | 90 tests | 30 tests |
|---|---|---|
| 96-well microplate | 6 units | 2 units |
| Buffer A (Extraction buffer) | 1 x 45ml | 1 x 15ml |
| Buffer B (Blocking solution) | 1 x 6ml | 1 x 2ml |
| Buffer C (Wash buffer) | 1 x 3ml | 1 x 1ml |
| Dipsticks | 1 x 90 units | 1 x 30 units |
| Tube 1 (DAB – 100X stock) | 1 x 300µl | 1 x 100µl |
| Tube 2 (reduced cytochrome c – 20X stock) | 1 x 1.5ml | 1 x 500µl |
| Tube 3 (Buffer for activity assay) | 1 x 30ml | 1 x 10ml |
Metabolism >> Pathways and Processes >> Mitochondrial Metabolism >> Cytochromes
Metabolism >> Pathways and Processes >> Mitochondrial Metabolism >> Mitochondrial markers
Kits/ Lysates/ Other >> Kits >> Cell Metabolism Kits >> Other Metabolism Assay
Signal Transduction >> Metabolism >> Mitochondrial
Our Abpromise guarantee covers the use of ab109878 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
FuncS
Functional Studies - Complex IV Rodent Enzyme Activity Dipstick Assay Kit - 90 tests (ab109878)

Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. 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.
ELISA - Complex IV Rodent Enzyme Activity Dipstick Assay Kit (ab109878)

Figure 1. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 100 µg of protein extract.
- Complex IV Rodent Enzyme Activity Dipstick Assay Kit (ab109878)

Figure 2. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Based on the standard curve, 50 µg of protein extract were loaded onto a dipstick for each sample. The figure shows three developed dipsticks from unknown samples (1-3). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 23-87% of normal Complex IV activity levels.
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Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. Dipstick ELISA Kits extend this concept by utilizing the well-established lateral flow concept, wherein capture antibodies are striped onto nitrocellulose membrane and a wicking pad draws the sample through the antibody bands. 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.

Figure 1. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 100 µg of protein extract.

Figure 2. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Based on the standard curve, 50 µg of protein extract were loaded onto a dipstick for each sample. The figure shows three developed dipsticks from unknown samples (1-3). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 23-87% of normal Complex IV activity levels.
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