Rhodamine B NHS ester (N-Hydroxysuccinimidyl rhodamine B ester) is an amine-reactive fluorescent probe (λex/λem: 564/589 nm, respectively). Rhodamine B NHS ester could react with Glycine [1].
In Vitro, Guide (The following are our recommended procedures. This guideline is for informational purposes only and should be modified according to your specific needs)[1]. 1. Preparation of working solution 1.1 Prepare 5 mL of Acetonitrile solution containing 0.4 mM Rhodamine B NHS ester. 2. Specific steps 2.1 Mix 0.5 mL of 10 mM glycine solution with 4.5 mL of H3BO3 buffer (pH = 8.5). 2.2 Add 5 mL of acetonitrile solution containing 0.4 mM Rhodamine B NHS ester to the mixture. 2.3 The reaction is carried out at room temperature for 4 h, and then diluted to 25 mL with mobile phase solution. 2.4 The mixture is sonicated for 3 min and centrifuged at 2500 rpm for 10 min for separation. 2.5 The clear supernatant is filtered through a 0.45 μm membrane filter and then subjected to LC-MS analysis. 2.6 The mobile phase is methanol-water-acetic acid solution (70:29.4:0.6, v/v), with a flow rate of 1 mL/min and a temperature of 25 °C.
References:
[1]. Meng Q, et al. Synthesis and application of N-hydroxysuccinimidyl rhodamine B ester as an amine-reactive fluorescent probe. Dyes and pigments, 2007, 73(2): 254-260.
In Vitro, Guide (The following are our recommended procedures. This guideline is for informational purposes only and should be modified according to your specific needs)[1]. 1. Preparation of working solution 1.1 Prepare 5 mL of Acetonitrile solution containing 0.4 mM Rhodamine B NHS ester. 2. Specific steps 2.1 Mix 0.5 mL of 10 mM glycine solution with 4.5 mL of H3BO3 buffer (pH = 8.5). 2.2 Add 5 mL of acetonitrile solution containing 0.4 mM Rhodamine B NHS ester to the mixture. 2.3 The reaction is carried out at room temperature for 4 h, and then diluted to 25 mL with mobile phase solution. 2.4 The mixture is sonicated for 3 min and centrifuged at 2500 rpm for 10 min for separation. 2.5 The clear supernatant is filtered through a 0.45 μm membrane filter and then subjected to LC-MS analysis. 2.6 The mobile phase is methanol-water-acetic acid solution (70:29.4:0.6, v/v), with a flow rate of 1 mL/min and a temperature of 25 °C.
References:
[1]. Meng Q, et al. Synthesis and application of N-hydroxysuccinimidyl rhodamine B ester as an amine-reactive fluorescent probe. Dyes and pigments, 2007, 73(2): 254-260.