Protonex™ NIR 770 NHS ester

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	535.64
Solvent	DMSO

Spectral properties

Excitation (nm)	748
Emission (nm)	769

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure

Overview SDS Protocol

Molecular weight

535.64

Excitation (nm)

748

Emission (nm)

769

Protonex™ NIR 770 NHS ester is the only cell-permeable acidotropic pH dye commercially available for developing NIR pH probes. It is an excellent building block that can be readily used for developing site-specific pH sensors for monitoring pH fluctuation in a spatial environment. It can be readily conjugated to amino-containing biomolecules (such as antibodies, peptides and amino-modified oligos). It easily penetrates live cell membranes, thus can be used for monitoring pH changes in live cells. It is complimentary to CypHer7E that is cell-impermeable, and often used to study most classes of cell surface receptors which internalize into the endocytic pathway when activated by a ligand. Protonex™ NIR 770 NHS ester is a flexible, generic fluorophore that can be coupled to a receptor or epitope tag antibody and monitors neutral to acidic intracellular events. It is one of the very few acidotropic pH probes that exhibit NIR fluorescence.

Calculators

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Protonex™ NIR 770 NHS ester to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	186.693 µL	933.463 µL	1.867 mL	9.335 mL	18.669 mL
5 mM	37.339 µL	186.693 µL	373.385 µL	1.867 mL	3.734 mL
10 mM	18.669 µL	93.346 µL	186.693 µL	933.463 µL	1.867 mL

Molarity calculator

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Spectrum

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Spectral properties

Excitation (nm)	748
Emission (nm)	769

Images

Figure 1. Image of Hela cells stained with Protonex™ NIR 770 NHS ester. Hela cells were washed 3 times with HH buffer and then incubated with 10 µM of Protonex™ NIR 770 NHS ester in HH buffer for 30 min at 37°C. The cells were then washed 3 times with HH buffer and incubated with HH buffer or pH=5.0 and pH=8.0 buffer (Cat# 21235) containing 10 µM of nigericin for 5 min at 37°C. Images were acquired using a fluorescence microscope equipped with a Cy7 filter set.

References

View all 50 references: Citation Explorer

Fisetin-Mediated Perturbations of Membrane Permeability and Intracellular pH in Candida albicans.
Authors: Kim, Younhee
Journal: Journal of microbiology and biotechnology (2024): 1-10

Generalization of the Ratiometric Method to Extend pH Range Measurements of the BCECF Probe.
Authors: Tafech, Alaa and Beaujean, Céline and Usson, Yves and Stéphanou, Angélique
Journal: Biomolecules (2023)

mOrange2, a Genetically Encoded, pH Sensitive Fluorescent Protein, is an Alternative to BCECF-AM to Measure Intracellular pH to Determine NHE3 and DRA Activity.
Authors: Sarker, Rafiquel and Tse, Chung Ming and Lin, Ruxian and McNamara, George and Singh, Varsha and Donowitz, Mark
Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (2022): 39-49

Energy Metabolism and Intracellular pH Alteration in Neural Spheroids Carrying Down Syndrome.
Authors: Kashirina, Alena and Gavrina, Alena and Kryukov, Emil and Elagin, Vadim and Kolesova, Yuliya and Artyuhov, Alexander and Momotyuk, Ekaterina and Abdyyev, Vepa and Meshcheryakova, Natalia and Zagaynova, Elena and Dashinimaev, Erdem and Kashina, Aleksandra
Journal: Biomedicines (2021)

Plastidial transporters KEA1 and KEA2 at the inner envelope membrane adjust stromal pH in the dark.
Authors: Aranda Sicilia, María Nieves and Sánchez Romero, María Elena and Rodríguez Rosales, María Pilar and Venema, Kees
Journal: The New phytologist (2021): 2080-2090

Effects of Diluent pH on Enrichment and Performance of Dairy Goat X/Y Sperm.
Authors: He, Qifu and Wu, Shenghui and Huang, Ming and Wang, Ying and Zhang, Kang and Kang, Jian and Zhang, Yong and Quan, Fusheng
Journal: Frontiers in cell and developmental biology (2021): 747722

Glomerular Mesangial Cell pH Homeostasis Mediates Mineralocorticoid Receptor-Induced Cell Proliferation.
Authors: Gekle, Michael and Mildenberger, Sigrid
Journal: Biomedicines (2021)

Intracellular pH regulation in mantle epithelial cells of the Pacific oyster, Crassostrea gigas.
Authors: Ramesh, Kirti and Hu, Marian Y and Melzner, Frank and Bleich, Markus and Himmerkus, Nina
Journal: Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology (2020): 691-700

Functional effects of urotensin-II on intracellular pH regulators in human radial artery smooth muscle cells.
Authors: Loh, Shih-Hurng and Chang, Chung-Yi and Huang, Shu-Fu and Chao, Shih-Chi and Lin, Wei-Kuo and Huang, Eagle Yi-Kung and Tsai, Chien-Sung and Tsai, Yi-Ting
Journal: Peptides (2020): 170236

Altered Degranulation and pH of Neutrophil Phagosomes Impacts Antimicrobial Efficiency in Cystic Fibrosis.
Authors: Hayes, Elaine and Murphy, Mark P and Pohl, Kerstin and Browne, Niall and McQuillan, Karen and Saw, Le Er and Foley, Clare and Gargoum, Fatma and McElvaney, Oliver J and Hawkins, Padraig and Gunaratnam, Cedric and McElvaney, Noel G and Reeves, Emer P
Journal: Frontiers in immunology (2020): 600033