SMCy5.5 

SMCy5.5 is a SMCy dye (ex. 638 nm; em. 705 nm). The high brightness of SMCy5.5 allows for effective tracking and imaging of lipid droplet exchange between adipocytes. SMCy5.5 can be used for lipid droplet labeling studies^[1].

Guidelines (The following is our recommended protocol. This protocol is provided as a guide only and should be modified according to your specific needs)^[1].
1) Cell seeding: One day before lipid droplet staining, seed KB cells at a density of 1.5×10⁶ cells in a T-25 culture flask.
2) Pre-staining treatment: The next day, remove the culture medium and rinse the adherent cells twice with PBS.
3) Dye incubation: Incubate cells with SMCy3.5 or SMCy5.5 (prepared in OptiMEM containing 0.5% DMSO at a concentration of 1 μM) for 3 h at 37°C, 90% humidity, and 5% CO₂.
4) Dye removal: After incubation, discard the dye-containing medium, rinse KB cells twice with PBS, add growth medium to ensure complete removal of excess dye, and then rinse again with PBS.
5) Cell treatment: After trypsinization, the cells are centrifuged at 15,000 rpm for 3 minutes, and the pellet is resuspended in growth medium. The cell suspension is centrifuged again, and the pellet is resuspended in growth medium.
6) Cell mixing and plating: Equal amounts of the two cell suspensions are thoroughly mixed (total cell number approximately 6×10⁶) and then plated at a density of 1×10⁵ cells/well in an IBiDi chambered coverglass.
7) Nuclear staining: Prior to imaging, cells are incubated with Hoechst (5μg/mL) for 15 min to stain the nuclei.
Fixation and Washing: Cells are fixed with 4% paraformaldehyde for 5 minutes and then washed three times with HBSS.
8) Imaging: Imaging is performed 18 and 48 h after cell attachment.
9) Imaging Equipment and Parameters: A Nikon Ti-E inverted microscope is used; the microscope settings are as follows: Hoechst (excitation 405 nm, emission 510 nm), SMCy3.5 (excitation 532 nm, emission 600 nm), and SMCy5.5 (excitation 638 nm, emission 705 nm).
10) Image Recording and Processing: Images are recorded using NIS Elements software and processed using Icy software.

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

693.63

C₄₁H₄₆BF₂N₃O₄

2417507-09-4

Solid

Brown to black

CC1(C2=C(C=CC3=CC=CC=C32)N(/C1=C/C=C/C=C/C4=O[B+3]([F-])([F-])O=C(N(C5CCCCC5)C(N6C7CCCCC7)=O)[C-]4C6=O)CCCC#C)C

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from 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 (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Collot M, et al. Ultrabright and Fluorogenic Probes for Multicolor Imaging and Tracking of Lipid Droplets in Cells and Tissues. J Am Chem Soc. 2018 Apr 25;140(16):5401-5411.  [Content Brief]