APF |
Katalog-Nr.GC42825
One-Click Copy Product Info
|
APF ist eine Fluoreszenzsonde, die selektiv und dosisabhÄngig bestimmte Spezies unter den ROS nachweisen kann, die hochgradig resistent gegen Autoxidation sind.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 359010-70-1
Sample solution is provided at 25 µL, 10mM.
_1-3.$$$39978408@51.jpg');)
_1-9.$$$38538795@65.jpg');)
_1-9.$$$39112701@51.jpg');)
_1-7.$$$37316672@70.jpg');)
_366-372.$$$36198801@70.jpg');)
.$$$38565142@65.jpg');)
_1867-1883.$$$33248023@67.jpg');)
_eads6055.$$$39977546@45.jpg');)
_eadm9805.$$$40080571@45.jpg');)
_eadj3020.$$$39666821@45.jpg');)
_1-20.$$$39757301@28.jpg');)
_1-24.$$$38898113@28.jpg');)
_273-287.$$$36806353@46.jpg');)
_1-16.$$$37156877@44.jpg');)
_1-19.$$$37460805@44.jpg');)
_1291-1307.$$$34518654@46.jpg');)
APF is a novel ROS indicator that selectively and dose-dependently fluoresces upon binding to certain reactive oxygen species in solution and in cells with a maximum excitation/emission wavelength of 490/515 nm. APF has higher specificity and stability than HDCFDA. APF is cell membrane permeable, non-fluorescent, and reacts with reactive oxygen species, such as hydroxyl radical, peroxynitroso anion, or hypochlorite anion, to produce strong green fluorescence, which can be detected by fluorescence microscopy or flow cytometry. After reacting with hydroxyl radicals, peroxynitrite anion or hypochlorite anion, APF produces strong green fluorescence, which can be detected by fluorescence microscope, high throughput imager, fluorescence zymography or flow cytometry. At the same time, it can also be coupled with a variety of molecules to achieve specific labeling detection, which is widely used in the fields of biomarkers, cell imaging and fluorescence microscopy[1].
References:
[1]. Cohn C A, Pedigo C E, Hylton S N, et al. Evaluating the use of 3'-(p-Aminophenyl) fluorescein for determining the formation of highly reactive oxygen species in particle suspensions[J]. Geochemical Transactions, 2009, 10: 1-9.
The following program is for reference only, the actual use of the program needs to be improved according to your actual situation.
Ⅰ.Stock solution preparation
The cryopreserved 1mg powder was rewarmed at room temperature for at least 20min, after which 472μL of anhydrous DMF was added to form a 5mM storage solution. After sufficient dissolution and mixing, dispense according to the amount of each use, and store at -20℃ away from light, avoiding repeated freezing and thawing.
Note: APF is soluble in DMSO, but some data show that DMSO is a quencher of hydroxyl radicals, which interferes with detection and reduces sensitivity. Therefore, DMSO is not recommended as a solvent.
Ⅱ.Solution system testing
Before the formal experiment, take a tube of APF storage solution and place it at room temperature, so that it can be fully dissolved. Afterwards, dilute the solution with a suitable buffer (e.g. HHBS buffer or PBS buffer) to the desired working solution concentration. It is recommended that the starting concentration should be 1-10μM, and the actual concentration should be optimized according to your own experimental system or references.
Ⅲ. Cell System Assay
1. Prepare cell suspension or live cells on slides.
2. Dilute DMF stock solution into a suitable buffer, the recommended starting concentration range is 1-10μM.
3. Incubate the cells with diluted APF for 20-60 minutes at 37°C. Adherent cells can be incubated without trypsin digestion.
4. Wash the cells 2-3 times, remove the excess probe solution and replace it with fresh buffer or medium.
5. Select the appropriate fluorescence instrument and detect at Ex/Em: 490/515nm.
Note:
1. Bovine serum albumin (BSA) and phenol red will affect the fluorescence value and should be used with caution.
2. The fluorescence produced by APF is much weaker than other reactive oxygen probes (e.g. DCFH), which is determined by the characteristics of the probe itself.
References:
[1]. Flemmig J, Zschaler J, Remmler J, et al. The fluorescein-derived dye aminophenyl fluorescein is a suitable tool to detect hypobromous acid (HOBr)-producing activity in eosinophils[J]. Journal of Biological Chemistry, 2012, 287(33): 27913-27923.
[2]. Cohn C A, Pedigo C E, Hylton S N, et al. Evaluating the use of 3'-(p-Aminophenyl) fluorescein for determining the formation of highly reactive oxygen species in particle suspensions[J]. Geochemical Transactions, 2009, 10: 1-9.
| Cas No. | 359010-70-1 | SDF | |
| Canonical SMILES | Nc1ccc(cc1)Oc1ccc2c(c1)Oc1cc(O)ccc1C12OC(=O)c2ccccc12 | ||
| Formula | C26H17NO5 | M.Wt | 423.4 |
| Löslichkeit | DMF: 20 mg/ml,DMSO: 10 mg/ml,Ethanol: 10 mg/ml,Ethanol:PBS (pH 7.2)(1:3): .5 mg/ml | 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. |
||
| 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 | 2.3618 mL | 11.8092 mL | 23.6183 mL |
| 5 mM | 472.4 μL | 2.3618 mL | 4.7237 mL |
| 10 mM | 236.2 μL | 1.1809 mL | 2.3618 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 27 reference(s) in Google Scholar.)