Complex I Rodent Protein Quantity Dipstick Assay Kit (ab109875)

Overview

• Product nameComplex I Rodent Protein Quantity Dipstick Assay KitSee all Complex I kits ...
• Sample type
  Cell culture extracts, Tissue
• Assay typeQuantitative
• Species 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.

• Tested applicationsSandwich ELISA more details

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	90 tests	30 tests
  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
  Extraction Buffer	1 x 45ml	1 x 15ml
  Gold-conjugated antibody (dried in microplate wells)	1 x 90 tests	1 x 30 tests

• Research Areas
  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Other Metabolism Assay

  • Signal Transduction
  • Metabolism
  • Mitochondrial

Applications

Complex I Rodent Protein Quantity Dipstick Assay Kit images

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  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

Protocol Booklet