02 Oxidative Stress

-SulfoBiotics- Sulfide dibimane

-SulfoBiotics- Sulfide dibimane

Biosulfur Analysis

  • Product code
    SB15  -SulfoBiotics- Sulfide dibimane
  • CAS No.
    1392113-30-2
  • Chemical name
    Bis(2,5,6-trimethylpyrazolo[1,2-a]pyrazole-1,7-dione-3-ylmethyl)sulfide
  • MW
    C20H22N4O4S=414.48
Unit size --
10 nmol x 5 Please inquire distributors about price.

Discription

Recent years, hydrogen sulfide (H2S) has been shown to exhibit various physiological activities such as vasodilation, cytoprotection, insulin secretion, and neurotransmitter regulation. Further, it has been attracting attention as an important signaling molecule following nitric oxide (NO) and carbon monoxide (CO). Hydrogen sulfide, like NO and CO, is recognized as a gaseous molecule, however its pKa is about 7, and at physiological pH, about 80% of hydrogen sulfide exists in the form of hydrogen sulfide ion (HS-). In addition, hydrogen sulfide ions take on various bonding forms and structures in vivo, and the details of their mechanism of action are still unknown, and elucidation of the biological functions of sulfur, particularly hydrogen sulfide, is eagerly awaited.

Monobromobimane is a thiol-specific fluorescent labeling agent that produces two molecules of Bimane for each molecule of hydrogen sulfide (Sulfide dibimane). Sulfide dibimane can be separated by HPLC from bimane compounds in which monobromobimane reacts with thiol compounds such as glutathione and cysteine. In addition, since the generated compounds are fluorescent, high-sensitivity analysis by fluorescence detection is possible.

Developer Dojindo Molecular Technologies, Inc.

Manual

Technical info

Fig. 1 Reaction of Monobromobimane with Hydrogen Sulfide

 

Fig. 2 Example of calibration curve

Fig. 3 HPLC measurement example


Column: ODS-3
Mobile Phase: A) 0.1% TFA/H2O, B) 0.1% TFA/Acetonitrile (TFA: trifluoroacetic acid)
B conc.: 5%-35% (0-5 min), 35%-55% (5-16 min), 55%-70% (16-23 min)
Detection: Fluorescence (Ex: 475 nm, Em: 390 nm)
Flow Rate: 1 ml/min
Column Temp: 40°C


Example of use:HPLC measurement: References 2), 3), 5)

References

Open References

1) G. L. Newton, R. Dorian and R. C. Fahey, Anal. Biochem., 1981114, 383.
2) E. A. Wintner, T. L. Deckwerth, W. Langston, A. Bengtsson, D. Leviten, P. Hill, M. A. Insko, R. Dumpit, E. Vanden 
Ekart, C. F. Toombs and C. Szabo, Br. J. Pharmacology2010160, 941.
3) X. Shen, C. B. Pattillo, S. Pardue, S. C. Bir, R. Wang and C. G. Kevil, Free Radic. Biol. Med.201150, 1021

Q & A

Q

Stability of Sulfide dibimane.

A

Sulfide dibimane is unstable to light, so it should be stored in a light-shielded place after opening the aluminum lamination zip.
After dissolving in acetonitrile, keep the solution shielded from light by aluminum foil, etc., and use it for measurement within the same day.
The acetonitrile solution is stable for several hours in a light-shielded condition.

Q

What concentration of Sulfide dibimane should be prepared?

A

Add 100 µl of acetonitrile (for HPLC) per tube of the product and dissolve to obtain a concentration of 0.1 mmol/l.

Dilute this solution with an arbitrary amount of acetonitrile to prepare the Sulfide dibimane calibration solution.

For preparation method, please refer to the manual of this product.

Q

Is there an expiration date?

A

It is stable for six months after delivery in an unopened package.
Due to the charactor of this reagent, it cannot be stored after dissolution, so please use the solution within the same day after preparation

Handling and storage condition

Specification
Appearance: Colorless to slightly yellow solid
Purity (HPLC): ≧ 97.0 %
Dye content: 0.540~0.600 (370 nm)
Handling and storage condition
0-5°C, Protect from light
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