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Product name
Complex I Rodent Protein Quantity Dipstick Assay Kit
See all Complex I products (7) ...
Sample type
Cell culture extracts, Tissue
Assay type
Quantitative
Cross reactivity
Reacts with
Mouse, Rat
Product overview
Contains 30 or 90 dipsticks and necessary components to quantify the levels of the fully assembled Complex I enzyme complex from mouse and rat 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 our Complex I Enzyme Activity Dipstick Assay Kit (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.
After diluting the 10X Blocking Buffer to 2X, store at 4°C.
For long-term storage, all buffers can be stored at 4°C.
Tested applications
Sandwich ELISAmore details
Properties
Storage instructions
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 |
Applications
Show applications key
Our Abpromise guarantee covers the use of ab109875 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
sELISA
Complex I Rodent Protein Quantity Dipstick Assay Kit images:
Sandwich ELISA - Complex I Rodent Protein Quantity Dipstick Assay Kit (ab109875)
An example using MS133 to quantify Complex I levels using various concentration of mouse brain tissue. Shown is a 1:2 dilution series using a positive control sample. Approximately 6 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 20 µg of mouse brain tissue extract. A one-site hyperbola line was generated for best-fit analysis using GraphPad.
Sandwich ELISA - Complex I Rodent Protein Quantity Dipstick Assay Kit (ab109875)
An example using MS133 to quantify Complex I levels using various concentration of mouse brain tissue. Based on the above results, 13 µg of tissue extract was loaded to determine intra-assay reproducibility. (Note: for a statistical analysis it is preferred to use two dipsticks for each sample; intra-assay CV's are typically <=10%). The average of the signal intensities was determined, and then the amount (µg) of Complex I was interpolated off of the standard curve.
Sandwich ELISA - Complex I Rodent Protein Quantity Dipstick Assay Kit (ab109875)
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.
Protocols
References for Complex I Rodent Protein Quantity Dipstick Assay Kit (ab109875)
ab109875 has not yet been referenced specifically in any publications.
Publishing research using ab109875? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"