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Oxygen Assays

Diogenes

Diogenes

$240.00

  • Cellular Luminescence Enhancement System for Superoxide Detection
  • Easy to Use – Specific for Superoxide
  • From 100-Fold to 600-Fold Enhancement
  • Requires Fewer Cells – Non-Cytotoxic

Catalog Number: CL-202

 
SKU: CL-202 Categories: ,

Description

Catalog Number: CL-202
  • Cellular Luminescence Enhancement System for Superoxide Detection
  • Easy to Use – Specific for Superoxide
  • From 100-Fold to 600-Fold Enhancement
  • Requires Fewer Cells – Non-Cytotoxic

Diogenes Cellular Luminescence Enhancement System is a superoxide chemiluminescent enhancer that is non-denaturing to living cells. Superoxide radical (O2–) is produced intracellularly as a consequence of aerobic metabolism and extracellularly by leukocytes in response to infection. The extent of oxidative burst produced by white blood cells (WBCs) when stimulated by f-met-leu-phe, phorbol esters, anti-Fc receptor antibodies, or LPS is a partial indicator of the immunocompetence of the cells tested.

Currently, the production of O2– by leukocytes is monitored by such cumbersome and indirect methods as measuring oxygen uptake in a Clark electrode (both in the presence and absence of cyanide) or measuring spectral changes caused by the reduction of cytochrome c. As a non-cytotoxic intermediate in the mechanism of photon production, Diogenes is ideally suited to the detection of cell-mediated superoxide production. The intensity of light produced by Diogenes in the presence of superoxide is directly proportional to the O2– concentration but is much higher than that achieved by using luminol. Therefore, Diogenes is ideal for monitoring cellular immunocompetence, utilizing a luminometer to quantify the light output. Any stimulant that activates an oxidase to produce extracellular superoxide is usable with Diogenes. Such means can be physiological or mimetic of the physiologic pathway.

 

Diogenes References:

Tsuyoshi Yamazaki, Chikage Kawai, […], and Futoshi Kuribayashi
Trop Med Health. 2011 June; 39(2): 41–45.

Miklós Geiszt, Jeffrey B. Kopp, […], and Thomas L. Leto
Proc Natl Acad Sci U S A. 2000 July 5; 97(14): 8010–8014.
Futoshi Kuribayashi1, Hiroyuki Nunoi, Kaori Wakamatsu, Shohko Tsunawaki, Kazuki Sato, Takashi Ito, and Hideki Sumimoto
EMBO Journal (2002) 21, 6312 – 6320 

Additional information

Weight 1 lbs
Dimensions 8 × 3 × 10 in

Protocol

Diogenes Protocol

Solution Preparation

  1. Add 1.0 ml of deionized water to the Diogenes Reagent that is contained in the vial. Mix until completely dissolved.
  2. Add 9.0 ml of deionized water to the Diogenes Activator that is contained in the bottle. Mix until completely dissolved.
  3. Add the contents in the vial (Diogenes Reagent) to the contents in the bottle (Diogenes Activator), recap the bottle and shake vigorously to mix.
  4. The combined solution (10 ml) comprises the Diogenes Complete Enhancer in its ready-to-use, final working strength, and will yield up to 100 assays.

Assay

  1. Add 5X104 – 5X105 cells contained in glucose media into a luminometer cuvette or microtiter plate.
  2. Add 100 μl of Diogenes Complete Enhancer Solution.
  3. Add 20 μl of stimulant (PMA, mAb197, fMLP, etc.).
  4. Read the results. The time to peak output will depend on the cell and stimulant used. The response of Diogenes to superoxide is instantaneous.

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Safety Overview

Diogenes Activator Safety Overview

Safety Summary (see SDS for complete information before using the product):

Appearance and Odor
Crystalline solid.

EMERGENCY OVERVIEW – IMMEDIATE HAZARD
CAUTION! MAY BE HARMFUL IF SWALLOWED.

EMERGENCY OVERVIEW – CHRONIC HAZARD WARNING
INGESTION MAY SEQUESTER CALCIUM AND CAUSE CALCIUM PHOSPHATE DEPOSITS IN THE KIDNEYS.

Diogenes Reagent Safety Overview

Safety Summary (see SDS for complete information before using product):

Toxic by inhalation, in contact with skin and if swallowed
Avoid contact with skin and eyes

EMERGENCY OVERVIEW – IMMEDIATE HAZARD
TOXIC BY INHALATION, IN CONTACT WITH SKIN AND IF SWALLOWED.

Full SDS

Diogenes Reagent

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Diogenes Activator

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OXYGEN ASSAY

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