Nitrocefin (Synonyms: Nitrocephin) |
رقم الكتالوجGC14823
One-Click Copy Product Info
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Nitrocefin هو ركيزة β-lactamase chromogenic التي تخضع لتغير لون مميز من الأصفر إلى الأحمر حيث يتم تحلل رابطة الأميد في حلقة β-lactam بواسطة β-lactamase
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 41906-86-9
Sample solution is provided at 25 µL, 10mM.
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Nitrocefin is a cephalosporin derivative with color-indicating properties [1]. When β-lactamase hydrolyzes the β-lactam ring of Nitrocefin, the compound changes color from yellow to red, enabling visual detection [2]. Nitrocefin is primarily used for rapid screening of bacteria for β-lactamase activity [3].
References:
[1]. Lee M, Hesek D, Mobashery S. A practical synthesis of nitrocefin[J]. The Journal of organic chemistry, 2005, 70(1): 367-369.
[2]. Worthington R J, Melander C. Overcoming resistance to β-lactam antibiotics[J]. The Journal of organic chemistry, 2013, 78(9): 4207-4213.
[3]. Chow C, Xu H, Blanchard J S. Kinetic characterization of hydrolysis of nitrocefin, cefoxitin, and meropenem by β-lactamase from Mycobacterium tuberculosis[J]. Biochemistry, 2013, 52(23): 4097-4104.
This protocol only provides experimental guidance. The actual application must be optimized and adjusted according to your research objectives, sample type, and detection platform [1-2].
1. Preparation of reagents and materials
Nitrocefin, DMSO, PBS, bacterial colonies, bacterial suspension or lysed cell samples, hydrolysis buffer.
2. Preparation of Nitrocefin solution
(1) Stock solution (1–10mg/mL): Dissolve 5–10mg Nitrocefin in DMSO, mix well, store in the dark, and store at –20℃ for several months.
(2) Working solution (0.5–1.0mg/mL): Dilute the stock solution in PBS or Assay Buffer to 0.5–1.0mg/mL. If the solution turns red (not yellow) after preparation, it means the concentration is too high and should be further diluted to a yellow solution. The working solution should be kept away from light, stored at -20℃, and used within 14 days or 2 months.
3. Rapid detection method for β-lactamase activity
(1) Slide test: Add one drop of working solution to a clean slide, inoculate a single colony and observe the appearance of red (within 20-30 minutes).
(2) Direct contact method: Drop the working solution on the colony and observe the color change to red within 20-30 minutes.
(3) Bacterial solution method: Add 3-5 drops of working solution to 1mL of bacterial solution. If the color changes to red within 20-30 minutes, it indicates a positive result.
(4) Cell lysis method: Ultrasonic lyse 1mL of bacterial solution and add 3-5 drops of working solution. If the color changes to red within 40 minutes, it indicates β-lactamase activity.
4. Photometric quantitative detection
(1) Standard solution and reaction system: Prepare the standard solution and standard gradient using Nitrocefin and Hydrolysis Buffer + DMSO.
(2) Sample processing: After centrifugation, resuspending, and ultrasonic lysis of the solid strain, the supernatant was collected for assay. Reaction mixture: Sample + Reaction Mix (e.g., Assay Buffer + Nitrocefin) was added to each well.
(3) Reading and calculation: Kinetic reading was performed at 490nm. It is recommended to read every minute for 30–60min. β-lactamase activity was calculated based on the standard curve and time changes.
(4) The unit is defined as the amount of enzyme that hydrolyzes 1µmol Nitrocefin per minute.
Operation precautions
(1) Avoid light during operation to prevent drug degradation.
(2) Do not use the working solution before it turns red to maintain the accuracy of the yellow formula.
(3) Experimental conditions should be optimized by gradient concentration and time for each sample type.
References:
[1]. Shannon K, Phillips I. ?-Lactamase detection by three simple methods: Intralactam, nitrocefin and acidimetric[J]. Journal of Antimicrobial Chemotherapy, 1980, 6(5): 617-621.
[2]. Allen C, Turner C, Kalsi S, et al. Development of a rapid phenotypic test on a microfluidic device for carbapenemase detection using the chromogenic compound nitrocefin[J]. Diagnostic Microbiology and Infectious Disease, 2020, 96(4): 114926.
| Cas No. | 41906-86-9 | SDF | |
| المرادفات | Nitrocephin | ||
| Chemical Name | (6R,7R)-3-((E)-2,4-dinitrostyryl)-8-oxo-7-(2-(thiophen-2-yl)acetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid | ||
| Canonical SMILES | [H][C@]1([C@@H]2NC(CC3=CC=CS3)=O)SCC(/C=C/C4=CC=C([N+]([O-])=O)C=C4[N+]([O-])=O)=C(C(O)=O)N1C2=O | ||
| Formula | C21H16N4O8S2 | M.Wt | 516.5 |
| الذوبان | 20mg/ml in DMSO, 20mg/mL in DMF | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.9361 mL | 9.6805 mL | 19.3611 mL |
| 5 mM | 387.2 μL | 1.9361 mL | 3.8722 mL |
| 10 mM | 193.6 μL | 968.1 μL | 1.9361 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >90.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 3 reference(s) in Google Scholar.)