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AB228552

Intracellular pH Assay Kit

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(4 Publications)

The Intracellular pH Assay Kit (ab228552) uses a proprietary fluorescent indicator for measuring relative intracellular pH changes.
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Functional Studies - Intracellular pH Assay Kit (AB228552)
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Supplier Data

Functional Studies - Intracellular pH Assay Kit (AB228552)

Carbachol dose response in CHO-M1 cells.

CHO-M1cells were seeded overnight in 60,000 cells per 100 μL per well in a 96-well black wall/clear bottom Costar plate. The growth medium was replaced with 50 μL/well of RatioWorks™ BCFL, AM dyeloading solution for 37°C for 1 hour, follow by 15 minutes incubation with 5 μL/well of 220 mM NH4Cl. Carbachol (200μL/well) was added by FlexStation (Molecular Devices) to achieve the final indicated concentrations. The fluorescent signal was generated using Ex/Em = 490/535 nm (cut off at 525 nm).

Key facts

Detection method

Fluorescent

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based (quantitative)

Assay Platform

Microplate reader

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Product details

The Intracellular pH Assay Kit (ab228552) utilizes a proprietary fluorescent indicator for measuring relative intracellular pH changes. It is an homogeneous, kinetic, live-cell fluorescent assay that utilizes either a standard procedure or acid-load procedure. The standard protocol is designed for measuring the therapeutic targets of interest with a decrease in intracellular pH upon treatment. The 'Acid-Load' procedure is designed to measure the increase of intracellular pH associated with changes in cellular metabolism due to GPCR activation or growth factor activity. With the 'Acid-Load' procedure, ammonium chloride solution is added after the fluorescent pH dye is loaded into cells in a minimum volume. This 'acid-loading' step is followed by the addition of agonist in a relatively large volume (~4X) of buffer. The sudden volume change initiates an efflux of ammonia (NH3) from the cells causing a rapid decrease in intracellular pH, and thus a decrease in fluorescence signal. The effect of agonist on the subsequent recovery of intracellular pH is measured by the relative fluorescence signal increase.
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What's included?

{ "values": { "1000Test": { "sellingSize": "1000 Test", "publicAssetCode":"ab228552-1000Test", "assetComponentDetails": [ { "size":"1 x 1 Vial", "name":"BCFL, AM", "number":"AB228552-CMP04", "productcode":"" }, { "size":"1 x 10 mL", "name":"50 mM probenecid", "number":"AB228552-CMP01", "productcode":"" }, { "size":"1 x 100 mL", "name":"HHBS", "number":"AB228552-CMP02", "productcode":"" }, { "size":"1 x 10 mL", "name":"10X Pluronic F127 Plus", "number":"AB228552-CMP03", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
Multi
Storage information
Please refer to protocols
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Product protocols

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Target data

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Publications (4)

Recent publications for all applications. Explore the full list and refine your search

American journal of cancer research 13:4721-4733 PubMed37970357

2023

Inhibition of Na+/H+ exchanger (NHE) 7 by 5-(N-ethyl-N-isopropyl)-Amiloride displays anti-cancer activity in non-small cell lung cancer by disrupting cancer stem cell activity and downregulating PD-L1 expression.

Applications

Unspecified application

Species

Unspecified reactive species

Bing-Yen Wang,Huan-Ting Shen,Yu-Ling Lee,Peng-Ju Chien,Wen-Wei Chang

Cells 11: PubMed36359862

2022

Melatonin Treatment Triggers Metabolic and Intracellular pH Imbalance in Glioblastoma.

Applications

Unspecified application

Species

Unspecified reactive species

Beatriz I Fernandez-Gil,Andrea Otamendi-Lopez,Alexandra Bechtle,Carla A Vazquez-Ramos,Neda Qosja,Paola Suarez-Meade,Rachel Sarabia-Estrada,Mark E Jentoft,Hugo Guerrero-Cázares,Germaine Escames,Paula Schiapparelli,Alfredo Quiñones-Hinojosa

Biochemistry and biophysics reports 27:101054 PubMed34189282

2021

CFTR-mediated anion secretion in parathyroid hormone-treated Caco-2 cells is associated with PKA and PI3K phosphorylation but not intracellular pH changes or Na/K-ATPase abundance.

Applications

Unspecified application

Species

Unspecified reactive species

Rattana Chaimana,Jarinthorn Teerapornpuntakit,Walailak Jantarajit,Kornkamon Lertsuwan,Saowalak Krungchanuchat,Nattapon Panupinthu,Nateetip Krishnamra,Narattaphol Charoenphandhu

Oncotarget 12:859-872 PubMed33889306

2021

High and synergistic activity between mTORC1 and PLK1 inhibition in adenocarcinoma NSCLC.

Applications

Unspecified application

Species

Unspecified reactive species

Elodie Montaudon,Rania El Botty,Sophie Vacher,Olivier Déas,Adnan Naguez,Sophie Chateau-Joubert,Damien Treguer,Ludmilla de Plater,Leïla Zemoura,Fariba Némati,André Nicolas,Alain Chapelier,Alain Livartowski,Stefano Cairo,Catherine Daniel,Marie Brevet,Elisabetta Marangoni,Didier Meseure,Sergio Roman-Roman,Ivan Bieche,Nicolas Girard,Didier Decaudin
View all publications
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions
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Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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