DDAO |
| Catalog No.GC61840 |
DDAO es una prometedora sonda fluorescente roja de infrarrojo cercano (NIR) con longitud de onda de excitaciÓn ajustable (600-650 nm) y longitud de onda de emisiÓn larga (Λem = 656 nm). DDAO puede diseÑarse para la detecciÓn de las actividades de diferentes enzimas, como β-galactosidasa, sulfatasa, proteÍna fosfatasa 2A, carboxilesterasa 2, albÚmina humana y esterasas.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 118290-05-4
Sample solution is provided at 25 µL, 10mM.
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DDAO is a near-infrared (NIR) red fluorescent probe with tunable excitation wavelength (600-650nm) and long emission wavelength (λem=656nm) and a relatively low pKa (~5)[1]. DDAO can be used to detect the activities of different enzymes, such as β-galactosidase, sulfatase, protein phosphatase 2A, carboxylesterase 2, human albumin, and esterase. There is a linear relationship between protein phosphatase 2A (PP-2A) concentration and DDAO-induced fluorescence[2]. DDAO can be used to study the phagocytosis of cancer cells by macrophages[3]. DDAO can be used to study the proliferation of B cells[4].
References:
[1] Hou J, Qian M, Zhao H, et al. A near-infrared ratiometric/turn-on fluorescent probe for in vivo imaging of hydrogen peroxide in a murine model of acute inflammation[J]. Analytica Chimica Acta, 2018, 1024: 169-176.
[2] Leira F, Vieites J M, Vieytes M R, et al. Characterization of 9H-(1, 3-dichlor-9, 9-dimethylacridin-2-ona-7-yl)-phosphate (DDAO) as substrate of PP-2A in a fluorimetric microplate assay for diarrhetic shellfish toxins (DSP)[J]. Toxicon, 2000, 38(12): 1833-1844.
[3] Candas-Green D, Xie B, Huang J, et al. Dual blockade of CD47 and HER2 eliminates radioresistant breast cancer cells[J]. Nature communications, 2020, 11(1): 4591.
[4] Malinova D, Wasim L, Newman R, et al. Endophilin A2 regulates B‐cell endocytosis and is required for germinal center and humoral responses[J]. EMBO reports, 2021, 22(9): e51328.
This plan only provides a guide, please modify it to meet your specific needs.
1. Solution preparation
(1) Stock solution: Dissolve DDAO in DMSO to prepare a stock solution with a concentration of 5mM.
Note: After unused stock solution is aliquoted, store it at -20°C or -80°C in the dark to avoid repeated freezing and thawing.
(2) Working solution: Before the formal experiment, dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) to the required working concentration, such as 1μM.
Note: Please adjust the optimal working concentration according to the actual situation or refer to the literature to set the gradient concentration by yourself. The working solution must be prepared and used immediately.
2. Using DDAO to study the phagocytic effect of macrophages on cancer cells[1](from the literature, for reference only)
(1) 0.8×106 RAW264.5 cells or 1×106 THP1 cells were seeded into 6-well plates.
(2) After incubation for 24h, the RAW264.7 cells were treated with 0.1μg/mL lipopolysaccharide (LPS) for 24h to activate them. THP1 cells were differentiated by treatment with 40nM phorbol 12-myristate 13-acetate (PMA) for 48h.
(3) Activated macrophages were stained with DiO (Cat. No. GC19515) at a final concentration of 40nM in culture medium for 20min and then rinsed three times with culture medium.
(4) Cancer cells were labeled with 1μM DDAO (5mM stock, DMSO, stored at −20°C) in PBS at 37°C for 15min and then washed with PBS containing 1% FBS.
(5) DDAO-labeled target cells (1×106) were added to DiO-stained macrophages (Mφ) and incubated at 37°C in a final volume of 2mL for 2h.
(6) After incubation, Mφ and target cells were harvested by washing three times with EDTA-PBS and then digested with 0.25% trypsin.
(7) Flow cytometry was used to evaluate double-labeled cells (DIO+/DDAO+), which represent cancer cells engulfed by mature Mφ.
3. Using DDAO to study B cell proliferation[2](from the literature, for reference only)
(1) B cells were labeled with 1μM DDAO and cultured in complete RPMI supplemented with 4μg/mL CD40L, 100ng/mL BAFF, 1μg/mL anti-IgM or 2.5μg/mL CpG.
(2) After 3 days of culture, DDAO dilution (and mCherry expression in CRISPR-targeted B cells) was measured by flow cytometry.
References:
[1] Candas-Green D, Xie B, Huang J, et al. Dual blockade of CD47 and HER2 eliminates radioresistant breast cancer cells[J]. Nature communications, 2020, 11(1): 4591.
[2]Malinova D, Wasim L, Newman R, et al. Endophilin A2 regulates B?cell endocytosis and is required for germinal center and humoral responses[J]. EMBO reports, 2021, 22(9): e51328.
| Cas No. | 118290-05-4 | SDF | |
| Canonical SMILES | O=C1C(Cl)=C2C(C)(C)C3=C(C=CC(O)=C3)N=C2C=C1Cl | ||
| Formula | C15H11Cl2NO2 | M.Wt | 308.16 |
| Solubility | DMSO : 10.42 mg/mL (33.81 mM) | 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 | 3.2451 mL | 16.2253 mL | 32.4507 mL |
| 5 mM | 0.649 mL | 3.2451 mL | 6.4901 mL |
| 10 mM | 0.3245 mL | 1.6225 mL | 3.2451 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: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 38 reference(s) in Google Scholar.)
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