NTPAN-MI 

NTPAN-MI is a fluorescent probe (excitation wavelength: 405 nm). NTPAN-MI is selectively activated after labeling unfolded proteins with exposed thiols, thereby reporting the degree of protein homeostasis^[1][2].

Guidelines (The following is our recommended protocol. This protocol is for guidance only and should be modified to suit your specific needs)^[2].
1 Stock Solution:
1) Dissolve the powder in DMSO to prepare 2 mM NTPAN-MI.
Note: Store at -20°C for up to 6 months.
2 Working Solution:
1) Dissolve the 2 mM NTPAN-MI stock solution in cysteine-free culture medium (CFM) to prepare a 50 μM NTPAN-MI staining solution.
3 Staining Steps
1) Seed 0.5 × 10⁵ Neuro-2a cells per well of an 8-well μ-slide in complete DMEM and culture overnight at 37°C to achieve approximately 80% confluence.
2) Treat the experimental cells with the stress solution.
3) Remove the stress solution and gently wash the cells once with PBS to remove any residual solution.
4) Add pre-warmed 50 μM NTPAN-MI staining solution to each well of an 8-well μ-slide and incubate for 30 minutes.
5) Discard the NTPAN-MI staining solution. Gently rinse the cells once with PBS.
6) Incubate the cells with nuclear and ER staining solutions for 15 minutes.
7) Discard the nuclear and ER staining solutions.
8) Gently rinse the cells once with PBS. Fix the cells with fixation buffer for 5 minutes at room temperature.
9) Discard the fixation buffer and store the cells in PBS. Samples can be stored at 4°C for up to 2 weeks until data collection is complete.
10) Observation of unfolded proteins: Set the excitation/emission wavelength of the confocal microscope to 405/495-555 nm (NTPAN-MI).

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

462.54

C₂₉H₂₆N₄O₂

2411398-95-1

Solid

Yellow to orange

N#C/C(C1=CC=C(N2C(C=CC2=O)=O)C=C1)=C(C3=CC=C(N(C)C)C=C3)\C4=CC=C(N(C)C)C=C4

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]. Owyong TC, et al. A Molecular Chameleon for Mapping Subcellular Polarity in an Unfolded Proteome Environment. Angew Chem Int Ed Engl. 2020 Jun 15;59(25):10129-10135.  [Content Brief]

  [2]. Zhang S, et al. Measuring Cysteine Exposure in Unfolded Proteins with Tetraphenylethene Maleimide and its Analogs. Methods Mol Biol. 2022;2378:3-18.  [Content Brief]