OxiSelect™ Free Hydrogen Sulfide Gas Assay Kit

Catalog No. ABIN5680065

Quick Overview for OxiSelect™ Free Hydrogen Sulfide Gas Assay Kit (ABIN5680065)

Reactivity: Others

Detection Method: Colorimetric

Application: Immunoassay (IA)

Product Details

Purpose: Hydrogen Sulfide Assay Kit is a novel technique for measurement of free H2S gas emission.

Brand: OxiSelect™

Characteristics: ' Hydrogen Sulfide Assay Kit is a novel technique for measurement of free H2S gas emission. Liquid samples in a 96 well microplate release H2S gas into the air which is captured in a gel spot containing Ag+ ions on the underside of the microplate lid just above the sample. The resulting silver sulfide (Ag2S) produced inside the spot creates a brownish color that can be measured colorimetrically on a microplate spectrophotometer. Each kit provides sufficient reagents to perform up to 100 assays, including positive control and unknown samples.

Components:

1. Sodium Sulfide : One vial containing 100 mg of Na2S•9H2O
2. Polymer Matrix : Two 1.5 mL vials.
3. 20X Silver Probe Solution : One 150 μL vial.

Application Details

Reagent Preparation:

• Lid Coating Mix: Transfer the proper amount of Polymer Matrix to a microcentrifuge tube and dilute the 20X Silver Probe Solution 1:20 into the Polymer Matrix. For example add 45 μL of 20X Silver Probe Solution to 855 μL of Polymer Matrix and mix well. This Lid Coating Mix volume is enough for 10 lid spots. The Lid Coating Mix is stable for one day at room temperature in a tightly capped tube. Note: Prepare only enough for immediate use by scaling the above example proportionally.
• Sodium Sulfide Positive Control: Prepare a 10 mM solution of Sodium Sulfide by weighing out 5 milligrams of solid Sodium Sulfide and resuspending the solid at 2.4 mg/mL in deionized water. Then dilute the 10 mM Sodium Sulfide 1:50 into distilled water to a final concentration of 200 μM. For example, for 1 mL of Sodium Sulfide Positive Control, add 20 μL of 10 mM Sodium Sulfide to 980 μL of deionized water. While the 10 mM Sodium Sulfide solution is stable for up to 4 hours in a tightly capped tube, the 200 μM Sodium Sulfide Positive Control should be prepared immediately before use in the assay. 5 Preparation of Coated Lid Notes:
• Make sure that the coated circular areas will cover the wells of interest when the lid is properly applied to the microplate.
• To avoid cross contamination of H2S gas between experimental wells, always arrange the experiment in the 96 well microplate so that a well containing a sample is bordered on every side by a blank well. 1. Place a 96 well lid upside down on a lab bench, or in a hood. 2. Draw up 30 μL of Lid Coating Mix (see Preparation of Reagents) with a micropipette and slowly apply about half of the Lid Coating Mix to the center of the circular area that covers one well. Use the pipette tip to evenly spread the Lid Coating Mix throughout the entire circular area. Then apply all but approximately 1-2 μL of the Lid Coating Mix remaining in the pipette tip to the circular area. Note: Leaving about 1-2 μL in the pipette tip ensures that no bubbles will be introduced into the coated circular area. If bubbles are accidently introduced into the circular area, use the pipette tip to gently aspirate away the bubbles. 3. Allow the inverted lid to dry undisturbed for 1 hour at room temperature. Do not touch or move the lid during this time.

Assay Procedure:

1. Add 300 μL of unknown sample, Sodium Sulfide positive control, or Negative Control to the wells of a 96 well microplate.
2. Cover the 96 well microplate with the Coated Lid (See Preparation of Coated Lid).
3. Incubate plate at room temperature undisturbed for one hour.
4. Transfer the Coated Lid to a new empty 96 well microplate. 6
5. Read absorbance of each microwell on a spectrophotometer using 405 nm as the primary wavelength.

Restrictions: For Research Use only

Handling

Storage: RT/4 °C

Storage Comment: Upon receipt, store the Sodium Sulfide at 4°C. Store all other kit components at room temperature.

Target Details

Background: Hydrogen sulfide (H2S) is a colorless gas containing a foul odor reminiscent of rotten eggs. High levels of H2S are corrosive, flammable and poisonous. H2S can be generated from the breakdown of organic matter by microbes when oxygen is not present (typically in swamps and sewers) by a process called anaerobic digestion. H2S can also be found in natural gas, emissions from volcanoes, and in water from some wells. The human body produces minute levels of H2S where the molecule functions as a gaseous messenger like carbon monoxide or nitric oxide. H2S acts as a cerebrovascular dilator and is produced by the enzyme cystathione gamma lyase. Through activation of potassium ATP channels, H2S can dilate arterioles. Furthermore, H2S stimulates vascular smooth cell apoptosis and blocks proliferation- based vascular remodeling. Finally, H2S has been shown to have neuroprotective and cardioprotective properties, lowering oxidative stress by increasing levels of glutathione by enhancing cystine and cysteine transport to neurons.