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.