Dabsyl chloride

Manufactured by Merck Group

Sourced in United States

Dabsyl chloride is a chromophoric labeling reagent used for the detection and quantification of amino acids, peptides, and proteins in various analytical techniques. It functions as a derivatizing agent, enabling the visualization and detection of the labeled compounds.

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5 protocols using dabsyl chloride

Fluorescent Probes Synthesis and Cell Assay

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tris-n-butylphosphine (TBP, >99%), 1,8-diazabicyclo[5.4.0] undec-7-ene (DBU, 98%), Acetic acid (ACA, >99.5%), Sodium methoxide (Ca. 5 mol/L in methanol), Tetrahydrofuran (THF), trifluoroAcetic acid (TFA) were purchased in Energy chemical company (Shanghai, China). Propylene sulfide (PS, 98%), Nile Red (98%), dansyl chloride (>99%), dabsyl chloride (>99%), hydrogen peroxide (H₂O₂) were purchased from Sigma (St. Louis, USA). Rosup was purchased from Beyotime Institute of Biotechnology (Beijing, China). L929 cell line was purchased from the Culture Collections of the Chinese Academy of Science (Shanghai, China). Nude mice was purchased from the animal center of the Chinese Academy of Science (Shanghai, China).

Zhou Z., Wang Y, & Hu P. (2022). Oxidation-Responsive Micelles for Drug Release Monitoring and Bioimaging of Inflammation Based on FRET Effect in vitro and in vivo. International Journal of Nanomedicine, 17, 2447-2457.

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Separation and Detection of DAP Isomers

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Separation of DAP isomers was performed according to Richaud et al. [32 (link)]. Isolated PGN of T. forsythia, E. coli, and P. gingivalis were hydrolysed with 6 N HCl containing 0.2% thioglycolic acid at 110 °C for 16 h. Samples were dried using a nitrogen evaporator and washed with distilled water for three times. m-DAP and LL-DAP were purchased from Sigma and used as standards. Dabsylation was performed by the method of Chang et al. [31 (link)], using 100 μg of samples or standards dissolved in 100 μl 0.1 M sodium bicarbonate buffer, pH 9.0. 200 μl of dabsyl chloride (4 nmol μl^− 1; Sigma) were added and samples were incubated at 70 °C for 20 min. Dried preparations were dissolved in 100–500 μl ethanol (70%, v/v) and 20 μl were injected onto a reversed-phase HPLC column (Ultimate 3000, C18, 150 × 4.6 mm). An isocratic elution was done at 37 °C with 12 mM ammonium phosphate, pH 6.5-acetonitrile-dimethylformamide (69:27:4, vol/vol/vol) at a flow rate of 0.6 ml min^− 1 and detection was done at 436 nm.

Mayer V.M., Hottmann I., Figl R., Altmann F., Mayer C, & Schäffer C. (2019). Peptidoglycan-type analysis of the N-acetylmuramic acid auxotrophic oral pathogen Tannerella forsythia and reclassification of the peptidoglycan-type of Porphyromonas gingivalis. BMC Microbiology, 19, 200.

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Soybean GABA Extraction and Analysis

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Soybean seeds (cultivar Yunhe, obtained from Jilin Province of China in 2012) were stored in polyethylene containers at −20°C. Standard samples of GABA (99% in purity), AG, EGTA, and dimethylaminoazobenzene sulfonyl chloride (dabsyl chloride, 99% in purity) were purchased from Sigma Chemical Co. Ltd. (St Louis, MO, USA). Acetonitrile was high-performance liquid chromatography (HPLC) grade. Other chemicals and reagents were of analytical grade.

Yang R., Guo Y., Wang S, & Gu Z. (2014). Ca2+ and aminoguanidine on γ-aminobutyric acid accumulation in germinating soybean under hypoxia–NaCl stress. Journal of Food and Drug Analysis, 23(2), 287-293.

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Synthesis and Characterization of Functional Oligomers

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Hexylamine (CAS 111-26-2), triethylamine (CAS 121-44-8), dabsyl chloride (CAS 56512-49-3), cholesterol (CAS 57-88-5), bilirubin (CAS 635-65-4), glucose (CAS 50-99-7), sucrose (CAS 57-50-1), phosphate (CAS 4265-44-2), and creatinine (CAS 60-27-5) were purchased from Sigma Aldrich, Bengaluru, India. NaCl (CAS 7647-14-5), KCl (CAS 7447-40-7) and mono-6-O-(p-toluenesulfonyl)-β-cyclodextrin (CAS 67217-55-4) were purchased from TCI chemicals, Dehradun, India. Starting materials for the functional oligomer synthesis, including OFP-Br and β-cyclodextrin functionalized with a thiol group (β-CD-SH), were prepared according to the reported procedures [46 (link),47 (link)]. Solvents were purchased from Merck, Darmstadt, Germany, and used without further purification.

Joshi V., Hussain S., Dua S., Arora N., Mir S.H., Rydzek G, & Senthilkumar T. (2022). Oligomer Sensor Nanoarchitectonics for “Turn-On” Fluorescence Detection of Cholesterol at the Nanomolar Level. Molecules, 27(9), 2856.

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Quantification of GABA in Fermented Milk

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Fermented milk was centrifuged and filtered through a Millex-HA 0.22-μm pore size filter to collect the supernatant. The concentration of GABA in the supernatant was analyzed using HPLC according to the dabsyl chloride (Sigma-Aldrich) derivatization method (Syu et al., 2008) . The GABA reagent standard was purchased from Sigma-Aldrich.
Briefly, the determination of dabsyl-GABA was carried out by HPLC (Agilent 1200, Santa Clara, CA).
A reversed-phase column (Agilent ZORBAX SB-C18, 250 mm × 4 mm, 5 μm particle size) coupled with a C18 cartridge was used. The column temperature was maintained at 35°C. The detection wavelength was 425 nm. The composition of the optimized mobile phase was methanol/CH 3 COONa solution (0.045 mol/L, pH 4) 65:35. The mobile phase was filtered through a 0.22μm filter membrane and degassed before use.

Han M., Liao W.Y., Wu S.M., Gong X, & Bai C. (2020). Use of Streptococcus thermophilus for the in situ production of γ-aminobutyric acid-enriched fermented milk. Journal of dairy science, 103(1).

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