Usage of Cefnitrothiophene:
1. Preparation of Cefnitrothiophene Storage Solution (10mg/ml)
(1) Before use, remove the product from the refrigerator and place it at room temperature for at least 20 minutes, followed by low-speed centrifugation
(2) Suck 100pu | high-quality DMSO and add 1mg of Cefnitrothiophene, fully dissolve and mix well. It can be packaged according to a single dosage and stored in dark at -20C.
2. Preparation of Cefnitrothiophene Working Solution (1mg/ml)
The typical working concentration of Cefnitrothiophene is 0.5-1.0mg/ml, and this preparation is generally yellow in color. Take 20ul of Cefnitrothiophene (10mg/ml) storage solution and add it to 180ul of PBS buffer (0.1M neutral pH). After mixing, obtain Cefnitrothiophene (1mg/ml) working solution. Excess working fluid can be stored in a dark place at -20C and must be used up within 2 weeks.
3. Rapid detection and analysis
Once exposed to a lactam producing bacterium, the sample to be tested can generally produce a positive reaction quickly. It is usually recommended to have a reaction time of 20-30 minutes and set a negative control.
(1) Surface analysis of glass slides
Take a drop of Cefnitrothiophene working solution (1mg/ml) onto a clean glass slide surface; b) Use the sterile inoculation ring to pick up a solid culture clone and mix it with the liquid drop. c) If red color can be observed within 20-30min, it indicates the activity of lactam enzyme. (If necessary, add more drops of Cefnitrothiophene working solution to prevent drying)
(2) Direct contact analysis method
Directly suck a drop of Cefnitrothiophene working solution (1mg/ml) to the surface of isolated colonies b) If red color can be observed within 20-30min, it indicates that lactam enzyme is lingual.
(3) Analysis method for broth culture solution
Add 3-5 drops of Cefnitrothiophene working solution (1mg/ml) to 1 m broth culture solution; T) If red color can be observed within 20-30min, it indicates the activity of lactam enzyme.
(4) Cell lysate analysis method
Ultrasound lysis of 1m cell suspension b) Add 3-5 commercial cephalosporin thiophene working solution (1mg/ml) to the lysis suspension; b) If red color can be observed within 20-30min, it indicates the activity of lactam enzyme.
(5) Filter paper analysis method
Saturate a small piece of filter paper (~3 x 3cm) to absorb Cefnitrothiophene working solution (1mg/ml), and then place it in a clean culture dish; B) Jump out a separated clone and smear it on the impregnated filter paper. c) If red color can be observed within 20-30min, it indicates the activity of lactam enzyme.
Shanghai Qixin New Materials Technology Co., Ltd. was established in 2016 and is located in Shanghai, China. It is a high-tech enterprise dedicated to the development of new materials, process optimization, and the research and development of pharmaceutical intermediates and reagents. Its main businesses include 7-amino-3-chloromethyl-2-cephem-2-carboxylic acid p-methoxybenzyl ester hydrochloride (CAS: 113479-65-5), Perrin F, ParyleneF (VT-4), Cefnitrothiophene, Nitrocefin, and others. At the beginning of its establishment, the company regarded technological innovation and research and development capabilities as its core competitiveness, and was determined to provide customers with good products and technical services with price and technological advantages.
