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PIPES has two pKa values. One pKa (6.76 at 25°C) is near the physiological pH which makes it useful in cell culture work. Its effective buffering range is 6.1-7.5 at 25° C. The second pKa value is at 2.67 with a buffer range of from 1.5 - 3.5. PIPES has been documented minimizing lipid loss when buffering glutaraldehyde histology in plant and animal tissues. Fungal zoospore fixation for fluorescence microscopy and electron microscopy were optimized with a combination of glutaraldehyde and formaldehyde in PIPES buffer. It has a negligible capacity to bind divalent ions.

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