Complex I Enzyme Activity Dipstick Assay Kit

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Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (AB109720)

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.

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AbReview39461****

Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (AB109720)

ab109720 was used to quantify complex I activity in HCT116 cells. The kit was used as described in the manual.

Shortly : cells were scraped and lysed in extraction buffer. Cell fragments were removed bey centrifugation and protein concentration was measured by Bradford assay. Increasing amounts of protein (as shown in image) were applied to each well as standard curve. A standard flatbed scanner was used instead of a Dipstick-reader. Due to the low contrast of the resulting bands, the brightness and contrast of the image was adjusted afterwards. Developed for 45 minutes as recommended by the manual.

This image is courtesy of an Abreview submitted by Christian Marx

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Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (AB109720)

An example using ab109720 to measure Complex I activity in human fibroblast samples. Developed dipsticks from a 1 : 2 dilution series using a positive control sample and the associated standard curve. Starting material was 30 μg of fibroblast protein extract.

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Functional Studies - Complex I Enzyme Activity Dipstick Assay Kit (AB109720)

An example using ab109720 to measure Complex I activity in human fibroblast samples. Based on the standard curve, 15 μg of protein extract were loaded onto a dipstick for each sample. The figure shows four developed dipsticks, a control sample (1) and three unknowns (2-4). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 14-61% of normal Complex I activity levels.