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).

Guide (The following is the experimental plan we recommend. This plan serves only as a reference guide. The specific operations should be adjusted according to your actual needs.)
In-situ labeled sample processing, based on gel-based active protein analysis ^[2]. Let the cells be placed on ice to thaw, and add 200 μL of cold DPBS along with the protein inhibitor mixture to each sample.
2. Disrupt the cells and centrifuge at 100,000 × g for 45 minutes.
3. Take the supernatant (the soluble protein fraction), resuspend it in 200 μL of cold DPBS (with protease inhibitors).
4. Standardize the protein concentration of each sample to 1 mg/mL, with a volume of 43 μL.
5. Perform click chemical reactions 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, with a final volume of 50 μL.
6. Incubate at room temperature for 1 hour, then add 20 μL of 4X SDS loading buffer.
7. Add the sample (30 μL) and perform electrophoresis on a 10% SDS-PAGE gel. Then, image it 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]