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Products:Signal Transduction >> Metabolism >> Mitochondrial
MSCatalog No. MS431-30
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Read our guarantee »Complex IV Human Protein Quantity Dipstick Assay Kit
See all Complex IV products (10) ...
Cell culture extracts, Tissue
Quantitative
Reacts with
Cow, Human
ab109877(MS431) contains 30 or 90 dipsticks and necessary components to quantify the levels of the fully assembled 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.
Based on the immunologic sandwich assay, this kit utilizes two monoclonal antibodies (mAbs) specific to different antigens present on the intact COX enzyme complex. One antibody is immobilized on nitrocellulose membrane in a thin line perpendicular to the length of the dipstick while the other is gold-conjugated for a visual signal. When assembled COX is present in the sample, a red line appears on the dipstick at the site of the COX antibody line. The signal intensity is directly related to the level of COX enzyme complex in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.
Sandwich ELISAmore details
Please see Notes section
| Components | 90 tests | 30 tests |
|---|---|---|
| Buffer A (Extraction buffer) | 1 x 45ml | 1 x 15ml |
| Buffer B (10X Blocking solution) | 1 x 1.2ml | 1 x 0.4ml |
| Buffer C (Wash buffer) | 1 x 3ml | 1 x 1ml |
| Dipsticks | 1 x 90 units | 1 x 30 units |
| Gold-conjugated antibody (dried in microplate wells) | 1 x 90 tests | 1 x 30 tests |
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 ab109877 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
sELISA
ELISA - Complex IV Human Protein Quantity Dipstick Assay Kit (ab109877)

An example using ab109877 to quantify Complex IV levels using various concentration of human fibroblast extract. Shown is a 1:2 dilution series using a positive control sample. Approximately 7 to 8 dipsticks are suitable for covering the entire working range and the blank for background levels. In this example the dilution series starts with 10 µg of fibroblast extract. A one-site hyperbola line was generated for best-fit analysis using GraphPad.
ELISA - Complex IV Human Protein Quantity Dipstick Assay Kit (ab109877)

An example using ab109877 to quantify Complex IV levels using various concentration of human fibroblast extract. Based on the above standard curve values, 5 µg of protein extract were loaded to each dipstick for the four unknown samples (1-4). Using GraphPad software, the signal intensity from the standard curve was interpolated and the quantity of Complex I in samples 1-5 was determined. Based on the above analysis, the unknown samples had between 4% and 64% of Complex IV levels compared to the control.
Sandwich ELISA - Complex IV Human Protein Quantity Dipstick Assay Kit (ab109877)

Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted, and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. Multiplexing dipstick assays have multiple target protein lines. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurred.
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An example using ab109877 to quantify Complex IV levels using various concentration of human fibroblast extract. Shown is a 1:2 dilution series using a positive control sample. Approximately 7 to 8 dipsticks are suitable for covering the entire working range and the blank for background levels. In this example the dilution series starts with 10 µg of fibroblast extract. A one-site hyperbola line was generated for best-fit analysis using GraphPad.

An example using ab109877 to quantify Complex IV levels using various concentration of human fibroblast extract. Based on the above standard curve values, 5 µg of protein extract were loaded to each dipstick for the four unknown samples (1-4). Using GraphPad software, the signal intensity from the standard curve was interpolated and the quantity of Complex I in samples 1-5 was determined. Based on the above analysis, the unknown samples had between 4% and 64% of Complex IV levels compared to the control.

Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted, and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. Multiplexing dipstick assays have multiple target protein lines. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurred.
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