Rhodamine-N3 chloride 

Rhodamine-N3 chloride is an azide-rhodamine fluorescent dye that can be used to label biomolecules containing alkyne groups^[1][2]. Rhodamine-N3 chloride is a click chemistry reagent, it contains an Azide group and can undergo copper-catalyzed azide-alkyne cycloaddition reaction (CuAAc) with molecules containing Alkyne groups. It can also undergo strain-promoted alkyne-azide cycloaddition (SPAAC) reactions with molecules containing DBCO or BCN groups (Ex/Em = 544/576 nm).

Sample processing for in situ-labeled samples, gel-based activity-based protein profiling^[2].
Thaw the cell pellet on ice and add 200 μL of cold DPBS plus protease inhibitor cocktail to each sample.
Lyse the cells and centrifuge at 100,000 × g for 45 minutes.
Take the supernatant (soluble proteome) and resuspend in 200 μL cold DPBS (plus protease inhibitors).
Standardize the protein concentration of each sample to 1 mg/mL in a volume of 43 μL.
Click chemistry is performed on each sample using a solution with a final concentration of 25 μM Rhodamine-N3 chloride, 1 mM TCEP, 100 μM TBTA, and 1 mM CuSO4 in a final volume of 50 μL.
Incubate at room temperature for 1 hour before adding 20 μL 4X SDS loading buffer.
The samples (30 μL) are then loaded and electrophoresed on a 10% SDS-PAGE gel and imaged using a fluorescence scanner.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

847.44

C₄₄H₅₉ClN₈O₇

2363751-90-8

Solid

Brown to black

576

544

[N-]=[N+]=NCCOCCOCCOCCNC(CCC(N1CCN(CC1)C(C2=C(C3=C4C=CC(N(CC)CC)=CC4=[O+]C5=C3C=CC(N(CC)CC)=C5)C=CC=C2)=O)=O)=O.[Cl-]

Room temperature in continental US; may vary elsewhere.

4°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)

* 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.

• [1]. Shijiao Cai, et al. Glycyrrhizic Acid-Induced Differentiation Repressed Stemness in Hepatocellular Carcinoma by Targeting c-Jun N-Terminal Kinase 1. Front Oncol. 2020 Jan 9;9:1431.  [Content Brief]

  [2]. Bryan R Lanning, et al. A road map to evaluate the proteome-wide selectivity of covalent kinase inhibitors. Nat Chem Biol. 2014 Sep;10(9):760-767.  [Content Brief]