Complex IV Rodent Enzyme Activity Dipstick Assay Kit (ab109878)

Overview

• Product nameComplex IV Rodent Enzyme Activity Dipstick Assay KitSee all Complex IV kits ...
• Sample type
  Cell culture extracts, Tissue
• Assay typeQuantitative
• Species reactivity
  Reacts with: Mouse, Rat
  
• Product overview

  ab109878(MS432) contains 30 or 90 dipsticks and necessary components to quantify the activity of the cytochrome c oxidase enzyme complex from mouse and rat samples. The kit includes sufficient materials to generate a standard curve and evaluate several unknown samples.

  
  Using an antibody immobilized on the dipstick that specifically recognizes COX, the enzyme complex is immno-captured (i.e immuno-precipitated in active form) and enzyme activity is determined directly on the dipstick. Using a traditional method for determining COX enzyme activity by histo-chemical methods and in-gel activity assays, Di-amino benzidinetetrachloride (DAB) serves as the reporter of COX activity. The greater the signal of precipitated DAB, the greater the amount of COX enzyme complex. In addition, the reaction is cyanide sensitive. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.

• Notes

  Store dipsticks at room temperature in their provided container and out of direct sunlight.

  High humidity conditions should be avoided.

  Store Buffer A, B, and C at 4°C or at -20°C for long term storage.

  Store Tubes 1 and 2 at -20°C or at -80°C for long term storage; they can also be aliquoted upon receipt to prevent freeze/thaw cycles.

  Tube 3 can be stored at room temperature.

• Tested applicationsFunctional Studies more details

Properties

• Storage instructionsPlease refer to protocols.

• Components	90 tests	30 tests
  96-well microplate	6 units	2 units
  Buffer B (Blocking solution)	1 x 6ml	1 x 2ml
  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
  Tube 1 (DAB 100X stock)	1 x 300µl	1 x 100µl
  Tube 2 (reduced cytochrome c 20X stock)	1 x 1.5ml	1 x 500µl
  Tube 3 (Buffer for activity assay)	1 x 30ml	1 x 10ml

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

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Cytochromes

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

  • Signal Transduction
  • Metabolism
  • Mitochondrial

Applications

Complex IV Rodent Enzyme Activity Dipstick Assay Kit images

• 

  Functional Studies - Complex IV Rodent Enzyme Activity Dipstick Assay Kit - 90 tests (ab109878)
  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.
• 

  ELISA - Complex IV Rodent Enzyme Activity Dipstick Assay Kit (ab109878)
  Figure 1. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Developed dipsticks from a 1:2 dilution series using a positive control sample and the associated standard curve. Starting material was 100 µg of protein extract.
• 

  - Complex IV Rodent Enzyme Activity Dipstick Assay Kit (ab109878)
  Figure 2. An example using ab109878 to quantify Complex IV enzyme activity in mouse protein extracts Based on the standard curve, 50 µg of protein extract were loaded onto a dipstick for each sample. The figure shows three developed dipsticks from unknown samples (1-3). The analysis of the signal intensity and interpolation from the standard curve showed that the unknown samples have between 23-87% of normal Complex IV activity levels.

Protocols

Protocol Booklet