Complex I Human Protein Quantity Dipstick Assay Kit (ab109722)
Overview
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Product name
Complex I Human Protein Quantity Dipstick Assay Kit
See all Complex I kits -
Sample type
Cell culture extracts, Tissue -
Assay type
Sandwich (quantitative) -
Species reactivity
Reacts with: Cow, Human -
Product overview
ab109722 contains 48 dipsticks and necessary components to quantify the levels of the fully assembled Complex I enzyme complex in 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, the kit utilizes two monoclonal antibodies (mAbs) specific to different antigens present on the Complex I enzyme complex. One antibody is immobilized on the nitrocellulose membrane in a thin line perpendicular to the length of the dipstick, while the other is gold-conjugated and combined with the sample mix. The sample contents containing the gold-conjugated mAb wick past the mAb immobilized on the dipstick. When assembled Complex I is present in the sample, a red line appears at the site of the anti-Complex I antibody line. The signal intensity is directly related to the amount of Complex I in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system. To identify defects in Complex I that do not affect enzyme assembly combine this assay with ab109720 to determine the relative specific activity of Complex I. -
Notes
All components are stable in their provided containers at room temperature out of direct sunlight.
After diluting the 10X Blocking Buffer to 2X, store at 4°C.
For long-term storage, all buffers can be stored at 4°C.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Reagents
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 48 tests Buffer B (10X Blocking solution) 1 x 0.4ml Dipsticks 1 x 30 units Extraction Buffer (ab260490) 1 x 15ml Gold-conjugated antibody (dried in microplate wells) 1 x 30 tests Wash buffer 1 x 2ml -
Research areas
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Alternative names
- NADH dehydrogenase
Images
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Figure 2. An example using ab109722 to quantify Complex I levels using various concentration of human fibroblast extract.
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Figure 1. An example using ab109722 to quantify Complex I levels using various concentration of human fibroblast extract.
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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.