Fluorometric Intracellular pH Assay Kit
SIGMA/MAK150 - sufficient for 1000 fluorometric tests
Product Type: Chemical
Fluorometric Intracellular Ph Assay Kit, Cat. No. MAK150: Carbachol dose response in CHO-M1 cells. CHO-M1 cells were seeded overnight in 60,000 cells per 100 μL per well in a 96-well black wall/clear bottom plate. The growth medium was replaced with 50 μL/well of dye-loading solution for 37 °C for 1 hour, follow by 15 minutes incubation with 5 μL/well of 220 mM NH4Cl. Carbachol (200 μL/well) was added to achieve the final indicated concentrations.
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| detection method | fluorometric |
| relevant disease(s) | neurological disorders; genitourinary disorders; cancer |
| storage temp. | −20°C |
| usage | sufficient for 1000 fluorometric tests |
| Application: | Fluorometric Intracellular pH Assay Kit has been used to measure the intracellular pH. |
| Application: | This kit can be utilized to measure changes in intracellular pH in cells treated under various conditions. |
| Biochem/physiol Actions: | The Fluorometric Intracellular pH Assay Kit utilizes a proprietary cell-permeable fluorescent indicator, BCFL-AM, for measuring relative intracellular pH changes. BCFL-AM is a superior alternative to the popular ratiometric pH probe, 2′,7′-bis-(2-carboxyethyl |
| Features and Benefits: | Superior alternative to BCECF |
| General description: | Intracellular pH changes are implicated in diverse physiological and pathological processes, including cell proliferation, apoptosis, fertilization, malignancy, multidrug resistance, ion transport, lysosomal storage disorders, and Alzheimer′s disease. This kit can be utilized to measure decreases in intracellular pH in cells treated under various conditions using a simple standard protocol. Alternatively, an Acid-Load procedure can be used to measure the increase of intracellular pH associated with changes in cellular metabolism due to GPCR activation, growth factor activity, or other cellular stimulation. 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. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12161503 |