AF488 NHS ester |
Catalog No.GC59158
One-Click Copy Product Info
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AF488 NHS ester is an amine-specific fluorescent probe with optimal excitation wavelength of 488nm and the emission wavelength of 525nm.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1374019-99-4
Sample solution is provided at 25 µL, 10mM.
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AF488 NHS ester is an amine-specific fluorescent probe with optimal excitation wavelength of 488nm and the emission wavelength of 525nm[1]. AF488 NHS ester is pH-sensitive and emits fluorescence at low pH values, which can be used to detect the entry of nanoparticles into lysosomes with low pH levels[2]. AF488 NHS ester can be used to label viruses for tracking virus-cell interactions[3]. AF488 NHS ester can be used to prepare antibodies against Staphylococcus aureus(S. aureus), thereby establishing a S. aureus detection method based on fluorescence immunosorbent assay[4].
References:
[1] Kendall C G, Stockton A M, Leicht S, et al. Amine analysis using AlexaFluor 488 succinimidyl ester and capillary electrophoresis with laser‐induced fluorescence[J]. Journal of Analytical Methods in Chemistry, 2015, 2015(1): 368362.
[2] Vaughan H J, Est-Witte S, Dockery L T, et al. A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery[J]. Science and Technology of Advanced Materials, 2024, 25(1): 2351791.
[3] Zhang S L X, Tan H C, Hanson B J, et al. A simple method for Alexa Fluor dye labelling of dengue virus[J]. Journal of virological methods, 2010, 167(2): 172-177.
[4] Kim J K, Yun H Y, Kim J S, et al. Development of fluorescence-linked immunosorbent assay for rapid detection of Staphylococcus aureus[J]. Applied Microbiology and Biotechnology, 2024, 108(1): 2.
Detection method based on AF488 NHS ester
This protocol only provides a guideline, and should be modified according to your specific needs
1. Prepare dyeing solution
(1) Prepare stock solution: Prepare AF488 NHS ester stock solution with a concentration of 1mM in high-quality anhydrous DMSO. Notice: Aliquot the unused stock solution and store it in the dark at -20°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with a suitable buffer (such as PBS) to prepare a working solution with a concentration of 1μM. Please adjust the concentration of the working fluid according to the actual situation, and prepare it now to avoid repeated freezing and thawing. Notice: The working solution must be prepared fresh on the day of the experiment.
2. Protein Preparation
(1) Exchange protein into 0.1M NaHCO3 buffer (pH 8.3) using a spin filter or desalting column.
(2) Measure absorbance at 280nm and calculate concentration and mix AF488 NHS ester and protein at a 3:1 molar ratio (dye:protein) to achieve=0.8-1.0 dye per protein.
(3) Incubate 1h at room temperature in the dark with gentle agitation.
3. Quenching and detection
(1) Add 10µl 1M Tris pH 8.0 per 100µl reaction volume and incubate 15min.
(2) Remove unreacted dye by gel-filtration or spin desalting pre-equilibrated with PBS, and measure by flow cytometry.
4. Precautions:
(1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
(2) For your safety and health, please wear a lab coat and disposable gloves when operating
(3) AF488 NHS ester should be stored in the dark throughout the experiment.
(4) Keep reaction pH 8.0–8.5; lower pH slows coupling, higher pH increases hydrolysis
(5) This protocol and all reagents are for scientific research use only.
References:
[1] Wang J, Bhattacharyya J, Mastria E, et al. A quantitative study of the intracellular fate of pH-responsive doxorubicin-polypeptide nanoparticles[J]. Journal of Controlled Release, 2017, 260: 100-110.
| Cas No. | 1374019-99-4 | SDF | |
| Formula | C31H32N4O13S2 | M.Wt | 732.74 |
| Solubility | good in water, DMF, DMSO | Storage | Store at -20°C, protect from light |
| 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.3647 mL | 6.8237 mL | 13.6474 mL |
| 5 mM | 272.9 μL | 1.3647 mL | 2.7295 mL |
| 10 mM | 136.5 μL | 682.4 μL | 1.3647 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: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 11 reference(s) in Google Scholar.)