Cyanine5 azide chloride
Based on 1 Customer Validation
Cyanine5 azide chloride is a Fluorescent probe for cell-selective bacterial protein labeling, as well as a fluorescent dye for single-molecule imaging and dynamic tracking of intracellular organelles. Cyanine5 azide chloride can conjugate with the alkynyl side chain of propargylglycine incorporated into bacterial proteins by the engineered methionyl-tRNA synthetase variant PraRS via copper (I)-catalyzed azide-alkyne [3+2] cycloaddition reaction, thereby enabling in-gel fluorescent visualization of labeled proteins. Cyanine5 azide chloride can diffuse across cell membranes and undergo copper-free click reaction with DBCO-functionalized scAVD pre-bound to DBCO-functionalized biotin, forming fluorescent nanoprobes with enhanced brightness, photostability and localization accuracy, and exhibiting discrete photobleaching steps. The excitation wavelength of Cyanine5 azide chloride is Ex = 633 nm.
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- Pureté : 99.63%
- CAS No.: 1267539-32-1
- Formule: C35H45ClN6O
- Masse moléculaire:601.22
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Stockage:
-20°C, sealed storage, away from moisture and light
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Activité biologique
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Chemical Information
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Solvant et solubilité
- Pureté et documentation
- Références
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Help & FAQs
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Activité biologique
Description
In Vitro
Guide (The following is our recommended protocol. This protocol is for guidance only and should be modified according to your specific needs).
1. Stock Solution Preparation
1.1 Solvent: No specific solvent mentioned.
2. Working Solution Preparation
2.1 Diluent: Serum-free Dulbecco’s modified Eagle medium (DMEM)[2].
2.2 Working concentration: 1 μM[2].
2.3 Note: Adjust working solution concentration as needed; prepare fresh before use.
3. Staining Procedure
3.1 For purified His-tagged bacterial marker proteins from bacterial cell lysates[1]:
3.1.1 Incubation conditions: Incubate with 25 µM Cyanine5 azide, alongside 100 µM CuSO4, 500 µM THPTA, and 5000 µM sodium ascorbate for 30 min.
3.1.2 Washing steps: No washing steps are included in the labeling reaction.
3.2 For adherent HeLa cells[2]:
3.2.1 Incubation conditions: Pre-incubate cells with 1 μM DBCO-PEG4-Biotin at 37°C for 1 hour; incubate cells with 1 μM Cyanine5 azide chloride working solution at room temperature for 1 hour.
3.2.2 Washing steps: Wash cells with PBS three times after incubation.
4. Controls
4.1 Negative controls:
4.1.1 HeLa cells expressing mCherry (without scAVD) with DBCO-PEG4-Biotin pre-incubation.
4.1.2 HeLa cells expressing mCherry-scAVD without DBCO-PEG4-Biotin pre-incubation.
5. Detection & Analysis
5.1 Instrument type: Fluorescence scanner[1], epifluorescence microscope[2], confocal microscope[2].
5.2 Ex/Em wavelengths: 633 nm excitation, 680/30 nm emission[1]; excitation filter FF02-632/22, dichroic mirror FF660-Di02, emission filter FF01-692/40[2]; 633 nm laser for excitation[2].
5.3 Result analysis:
5.3.1 Fluorescence localization: Fluorescence is localized to purified bacterial marker proteins (GFPm) metabolically labeled with propargylglycine (Pra) via PraRS expression[1]; Cy5 dots localize to the cytosol in cytosol-targeted scAVD-overexpressing cells, or to endoplasmic reticulum (ER) membranes in ER-targeted scAVD-overexpressing cells[2].
5.3.2 Color changes: Red fluorescence is detected from Cyanine5 azide-labeled samples[1][2].
5.3.3 Fluorescence intensity and photobleaching: Cy5 dots formed in scAVD-overexpressing cells show higher brightness and longer emission duration before photobleaching compared to single Cy5-azide dye[2]; some Cy5 dots exhibit discrete four-step fluorescence intensity decrease during photobleaching, indicating four Cy5-azide dyes conjugated per scAVD molecule[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Emission (Em)
668
Chemical Information
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CAS No. 1267539-32-1
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Appearance Solid
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Masse moléculaire 601.22
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Formule C35H45ClN6O
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Color Light brown to brown
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SMILES
[N-]=[N+]=NCCCNC(CCCCCN1C2=C(C(C)(C)/C1=C\C=C\C=C\C3=[N+](C)C4=C(C3(C)C)C=CC=C4)C=CC=C2)=O.[Cl-]
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
-20°C, sealed storage, away from moisture and light
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)
Solvant et solubilité
In Vitro:
DMSO : 100 mg/mL (166.33 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Pureté et documentation
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Fiche technique (288 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Instruction de manipulation (2659 KB)
Références
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.6633 mL | 8.3164 mL | 16.6328 mL | 41.5821 mL |
| 5 mM | 0.3327 mL | 1.6633 mL | 3.3266 mL | 8.3164 mL | |
| 10 mM | 0.1663 mL | 0.8316 mL | 1.6633 mL | 4.1582 mL | |
| 15 mM | 0.1109 mL | 0.5544 mL | 1.1089 mL | 2.7721 mL | |
| 20 mM | 0.0832 mL | 0.4158 mL | 0.8316 mL | 2.0791 mL | |
| 25 mM | 0.0665 mL | 0.3327 mL | 0.6653 mL | 1.6633 mL | |
| 30 mM | 0.0554 mL | 0.2772 mL | 0.5544 mL | 1.3861 mL | |
| 40 mM | 0.0416 mL | 0.2079 mL | 0.4158 mL | 1.0396 mL | |
| 50 mM | 0.0333 mL | 0.1663 mL | 0.3327 mL | 0.8316 mL | |
| 60 mM | 0.0277 mL | 0.1386 mL | 0.2772 mL | 0.6930 mL | |
| 80 mM | 0.0208 mL | 0.1040 mL | 0.2079 mL | 0.5198 mL | |
| 100 mM | 0.0166 mL | 0.0832 mL | 0.1663 mL | 0.4158 mL |
Cyanine5 azide chloride Related Classifications
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