Acridinium NHS ester |
| Catalog No.GC42711 |
Acridinium NHS ester (AE NHS) is a protein and nucleic acid label at primary amines in biomolecules and produce chemiluminescence in the presence of hydrogen peroxide (H2O2)
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 177332-37-5
Sample solution is provided at 25 µL, 10mM.
Acridinium NHS ester (AE NHS) is a protein and nucleic acid label at primary amines in biomolecules and produce chemiluminescence in the presence of hydrogen peroxide (H2O2)[1] [2].
Acridinium NHS ester is applied in DNA labeling ex vitro(such as 2.95mmol/L, 30min, 37°C)[3], and protein labeling ex vitro (such as 10μM)[4]. Acridinium NHS ester–labeled antibody is also applied in microRNA detection[5], and viral genome of SARS-CoV-2[6].
References:
[1] Arnold, L J Jr et al. “Assay formats involving acridinium-ester-labeled DNA probes.” *Clinical chemistry* vol. 35,8 (1989): 1588-94.
[2] Natrajan, Anand, David Wen, and David Sharpe. "Synthesis and properties of chemiluminescent acridinium ester labels with fluorous tags." *Organic & Biomolecular Chemistry* 12.23 (2014): 3887-3901.
[3] He, Yi et al. “DNA sensor by using electrochemiluminescence of acridinium ester initiated by tripropylamine.” *Analytical and bioanalytical chemistry* vol. 399,10 (2011): 3451-8. doi:10.1007/s00216-010-4157-y
[4] Lai, Yongquan et al. “Using acridinium ester as the sonochemiluminescent probe for labeling of protein.” *The Analyst* vol. 134,1 (2009): 131-7. doi:10.1039/b813330a
[5] Kappel, Andreas et al. “MicroRNA in vitro diagnostics using immunoassay analyzers.” *Clinical chemistry* vol. 61,4 (2015): 600-7. doi:10.1373/clinchem.2014.232165
[6]Pham, Jimmykim et al. “Performance Characteristics of a High-Throughput Automated Transcription-Mediated Amplification Test for SARS-CoV-2 Detection.” *Journal of clinical microbiology* vol. 58,10 e01669-20. 22 Sep. 2020, doi:10.1128/JCM.01669-20
Protocol for labeling protein with Acridinium NHS ester [1]
1.Ultrasound decomposition of Acridinium NHS ester.
1.1Acridinium NHS ester test stock solution: Anhydrous DMF is used to dissolve Acridinium NHS ester, and the stock solution with a concentration of 1-5mg/ml is configured. After packaging, the stock solution is stored at -20℃ or -80℃ away from light.
1.2 Preparation of test solution: The test stock solution was diluted with a mixture of DMF and H2O with a volume ratio of 2:3 and configured into 8mL test solution containing 10μM (0.01mM) Acridinium NHS ester.
1.3 Irradiate acridinium ester solution by ultrasound for 15s, and then transfer 16ml 1mol/L sodium hydroxide to the solution followed by irradiation of ultrasound.
1.4 Irradiate with ultrasound lasted for 1min to ensure complete reaction.
2.Measurement of sonogenerated chemiluminescence (SCL).
2.1 All the SCL measurements performe under the conditions: ultrasound out-powers and frequency were 300W and 20kHz, respectively; the work voltage of the photomultiplier tube was 850V.
2.2 All SCL experiments are carried out in a glass cell fitted with an optically flat window.
2.3 The sonicated solution should be air-saturated and the volume of the sonicated solution was 8mL.
2.4 The filter with permeation of 440nm is used to decrease the intensity of background SL generated from solvent.
2.5 The change of solution temperature generated by the ultrasound only influenced slightly the SCL intensity, so there is no need to use a constant temperature device.
3.Preparation of labeled antibodies.
3.1 Label stock solution: Acridinium NHS ester stock solution is configured in DMSO, such as 0.5-2mM.
Note: Please adjust the concentration of stock liquid and working fluid according to the actual situation. Unused stock solution is recommended to be stored away from light at -20°C or -80°C after packaging.
3.2 Transfer a solution of 30mg antibody (anti-human insulin guinea pig polyclonal antibody in this case) in 50mL labeling buffer to a small brown plastic test tube, then add 5mL 0.5mM Acridinium NHS ester* (previously diluted in anhydrous DMF and stored at 20°C).
3.3 Stir the mixture at room temperature for 1.5h.
3.4 remove excess acridinium ester label by dialysis with 0.1mol/L NaH2PO3 (4°C; 10h).
3.5 Transfer the mixture tto a dialytic bag, and then put the dialytic bag into a glass beaker filled with 100mL dialysate buffer.
3.6 The purification is carried out at 4°C for 10h and repeated three times to remove the unconjugated Acridinium NHS ester from AE-conjugated polyclonal antibody completely.
3.7 Collect 50mL AE-conjugated and dilute to 500mL with pH 7.4 buffer (0.1M phosphate buffer, 0.15M NaCl) for further use.
References:
[1]Lai, Yongquan et al. “Using acridinium ester as the sonochemiluminescent probe for labeling of protein.” The Analyst vol. 134,1 (2009): 131-7. doi:10.1039/b813330a
| Cas No. | 177332-37-5 | SDF | |
| Canonical SMILES | C[N+]1=C2C(C=CC=C2)=C(C(OC3=CC=C(CCC(ON4C(CCC4=O)=O)=O)C=C3)=O)C5=C1C=CC=C5.FC(F)(S(=O)([O-])=O)F | ||
| Formula | C28H23N2O6•CF3SO3 | M.Wt | 632.6 |
| Solubility | Soluble in DMSO | 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 | 1.5808 mL | 7.9039 mL | 15.8078 mL |
| 5 mM | 0.3162 mL | 1.5808 mL | 3.1616 mL |
| 10 mM | 0.1581 mL | 0.7904 mL | 1.5808 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 ddH2O, 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: >97.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 8 reference(s) in Google Scholar.)
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