TMA-DPH (solution) 

TMA-DPH (solution) is a hydrophobic fluorescent membrane probe (Ex=355 nm; Em=430 nm). TMA-DPH is able to anchor on the cell surface and localize to different regions of the phospholipid bilayer. By analyzing the fluorescence polarization values of TMA-DPH in the plasma membrane and membrane substructures, the fluidity of the cell membrane can be determined^[1][2][3].
Solution Concentration: 10 mM

1. Solution preparation
1.1 Preparation of stock solution
Solvent: DMSO
Concentration: 10 mM (optimized according to the experiment).
1.2 Preparation of working solution
Dilute with PBS or serum-free medium (pH 7.4) (optimized according to the experiment). The corresponding stock solution can be diluted according to the actual situation. Note that if the solvent is DMSO, the cytotoxicity of DMSO must be considered, and a solvent control should be prepared; if the solvent is pure water, the working solution needs to be filtered and sterilized before adding cells.
Note: The working solution should be prepared and used immediately. Keep it away from light.

2. Cell staining (suspended cells)
2.1 Collect cells by centrifugation and wash twice with PBS for 5 minutes each time.
2.2 TMA-DPH (0.5-5 μM) in Hanks and 20 mM Hepes buffer pH 7.4, and incubate it at 37°C, for 5-30 minutes (optimized according to the experiment).
2.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
2.4 Add PBS to wash the cells twice, 5 minutes each time.
2.5 Wash the cells and resuspended in the appropriate Hepes buffer pH 7.4.
2.6 Observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture the adherent cells on a sterile coverslip.
3.2 Remove the coverslip from the culture medium and remove the excess culture medium.
3.3 Add TMA-DPH (0.5-5 μM) in Hanks and 20 mM Hepes buffer pH 7.4, and incubate it at 37°C, for 5-30 minutes (optimized according to the experiment).
3.4 Wash the cells and resuspended in the appropriate Hepes buffer pH 7.4.
3.5 Fluorescence microscopy detection (Ex/Em = 355/430 nm).
Note: If flow cytometry is required, the cells need to be digested with trypsin and resuspended before staining.

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

461.62

C₂₈H₃₁NO₃S

115534-33-3

C[N+](C)(C)C1=CC=C(/C=C/C=C/C=C/C2=CC=CC=C2)C=C1.[O-]S(=O)(C3=CC=C(C)C=C3)=O

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Illinger D, et al. The kinetic aspects of intracellular fluorescence labeling with TMA-DPH support the maturation model for endocytosis in L929 cells. J Cell Biol. 1994 May;125(4):783-94.  [Content Brief]

  [2]. Illinger D, et al. A comparison of the fluorescence properties of TMA-DPH as a probe for plasma membrane and for endocytic membrane. Biochim Biophys Acta. 1995 Oct 4;1239(1):58-66.  [Content Brief]

  [3]. Benedetti A, et al. Plasma membrane fluidity in isolated rat hepatocytes: comparative study using DPH and TMA-DPH as fluorescent probes. J Gastroenterol Hepatol. 1989 May-Jun;4(3):221-7.  [Content Brief]