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Spexyte™ Intracellular pH Calibration Buffer Kit
Intracellular pH (pHi) plays an important modulating role in many cellular events, including cell volume regulation, cellular metabolism, calcium regulation, receptor-mediated signal transduction, ion transport, endocytosis, and other cellular processes. Intracellular pH is generally 6.8-7.4 in the cytosol and 4.5-6.0 in the acidic organelles. Intracellular pH changes have significant physiological effects, e.g., the pH-dependent concentration of intracellular messengers such as Ca2+ and cAMP affects cellular signaling. Several recent reports showed the dysregulated pH is emerging as a hallmark of cancer cells. Spexyte™ Intracellular pH Calibration Buffer Kit provides a range of pH calibration buffers (pH 4.5-8.0) with nigericin, which modulate the intracellular pH with the external pH in the presence of 100-150 mM K+. When used in conjunction with pH indicators, such as BCFL, AM or BCECF, AM, Spexyte™ Intracellular pH Calibration Buffer Kit can create a standard curve which is used to determine the intracellular pH.
Standard curve created using BCFL, AM with Spexyte™ Intracellular pH Calibration Buffer Kit. Hela cells were incubated with 5µM BCFL, AM for 30 minutes at room temperature. The Intracellular pH Calibration Buffer Kit ( Cat#21235) was used to clamp the intracellular pH with extracellular buffers at pH 4.5 to 8.0. Intracellular pH vs. relative fluorescence ratio of Ex/Em= 440/ 530 nm and 500 nm/530 nm were plotted and a 4-parameter trendline was fitted to get the pH standard curve.
Standard curve created using BCFL, AM with Spexyte™ Intracellular pH Calibration Buffer Kit. Hela cells were incubated with 5µM BCFL, AM for 30 minutes at room temperature. The Intracellular pH Calibration Buffer Kit ( Cat#21235) was used to clamp the intracellular pH with extracellular buffers at pH 4.5 to 8.0. Intracellular pH vs. relative fluorescence ratio of Ex/Em= 440/ 530 nm and 500 nm/530 nm were plotted and a 4-parameter trendline was fitted to get the pH standard curve.
protocol
Protocol
sds
SDS
COA
CatalogSize
Price
Quantity
21235100 Tests
Price
 
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microscope
ExcitationTexas Red/FITC filter
EmissionTexas Red/FITC filter
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation440, 500 nm
Emission530 nm
Cutoff515 nm
Recommended plateBlack wall/clear bottom
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Page updated on October 8, 2025