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Dihydroethidium |
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Cat.No.£ºIC-016360
Images of Hela cells stained with the Dihydroethidium in a Costar black wall/clear bottom 96-well plate. A: Untreated control cells. B: Cells treated with 100 ¦ÌM tert-butyl hydroperoxide (TBHP) for 30min before staining. |
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Product Information |
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Biological Activity |
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Dihydroethidium(Hydroethidine; PD-MY 003) is a superoxide indicator; exhibits blue-fluorescence in the cytosol until oxidized, where it intercalates within the cell''''s DNA, staining its nucleus a bright fluorescent red. |
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Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines) |
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Species |
Mouse |
Rat |
Rabbit |
Guinea pig |
Hamster |
Dog |
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Weight (kg) |
0.02 |
0.15 |
1.8 |
0.4 |
0.08 |
10 |
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Body Surface Area (m2) |
0.007 |
0.025 |
0.15 |
0.05 |
0.02 |
0.5 |
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Km factor |
3 |
6 |
12 |
8 |
5 |
20 |
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Animal A (mg/kg) = Animal B (mg/kg) multiplied by (Animal B Km / Animal A Km) |
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Chemical Information |
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Molecular Weight |
315.41 |
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Formula |
C21H21N3 |
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CAS Number |
104821-25-2 |
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Purity |
>98% |
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Solubility |
¡Ý 50 mg/mL (158.52 mM) in DMSO |
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Storage |
at -4¡ãC,Protect |
from light |
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References Information |
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[1]. Zielonka J, et al. Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem. 2012 Jan 27;287(5):2984-95. |
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Order Information
Product Name
Cat.No.
Quantity
Price($)
Price($)
Availability
Dihydroethidium
IC-016360
5mg
$105.00
€87.50
In stock
Dihydroethidium
IC-016360
10mg
$195.00
€162.50
In stock
Dihydroethidium
IC-016360
50mg
$320.00
€266.67
In stock
SAMPLE EXPERIMENTAL PROTOCOL
1. Add equal volume (such as 100 ¦ÌL of the cells in growth medium) of the dye working solution to the cells, and incubate the cells at RT or 37¡ãC for 5 to 60 minutes.
2. Determine the baseline fluorescence intensity of a sample of the loaded cells prior to exposing the cells to experimental inducements.
3. Negative controls should be assessed as follows:
a. Examine the fluorescence of cell-free mixtures of dye and buffer/media with and without the inducer. In the absence of extracellular esterases and other oxidative enzymes, the gradual increase in fluorescence over time may be related to spontaneous hydrolysis, atmospheric oxidation, and/or light- induced oxidation.
b. Examine the fluorescence of untreated (control) loaded cells that have been maintained in growth medium or buffer. In healthy cells, oxygen radicals are eliminated by cellular enzymes and/or natural antioxidants. Following thedye-loading recovery period, healthy cells should exhibit a low level of fluorescence that is relatively stable for the duration of the experiment; however, a gradual increase (due to auto-oxidation) or decrease (due to loss of dye from cells or photobleaching) in fluorescence may be observed. In the absence of any stimulus or inducement, a burst of fluorescence in healthy, untreated cells could indicate progress to cell death or some other oxidative event.
4. Positive controls may be stimulated with tert-butyl hydroperoxide (TBHP) to a final concentration of ~100 ¦ÌM (increase or decrease dose based on the sensitivity and response of the cells).
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