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Hardonk MJ, Haarsma TJ, Dijkhuis FW, et al. Influence of fixation and buffer treatment on the release of enzymes from the plasma membrane. Histochemistry. 1977;54(1):57-66doi: 10.1007/BF00493329.
Hardonk, M. J., Haarsma, T. J., Dijkhuis, F. W., Poel, M., & Koudstaal, J. (1977). Influence of fixation and buffer treatment on the release of enzymes from the plasma membrane. Histochemistry, 54(1), 57-66. https://doi.org/10.1007/BF00493329
Hardonk, M J, et al. "Influence of fixation and buffer treatment on the release of enzymes from the plasma membrane." Histochemistry vol. 54,1 (1977): 57-66. doi: https://doi.org/10.1007/BF00493329
Hardonk MJ, Haarsma TJ, Dijkhuis FW, Poel M, Koudstaal J. Influence of fixation and buffer treatment on the release of enzymes from the plasma membrane. Histochemistry. 1977 Oct 03;54(1):57-66. doi: 10.1007/BF00493329. PMID: 144099.
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Histochemistry. 1977 Oct 03;54(1):57-66. doi: 10.1007/BF00493329.

Influence of fixation and buffer treatment on the release of enzymes from the plasma membrane.

Histochemistry

M J Hardonk, T J Haarsma, F W Dijkhuis, M Poel, J Koudstaal

PMID: 144099 DOI: 10.1007/BF00493329

Abstract

1. Pretreatment of frozon cryostat sections with formaldehyde or calcium ions inhibits diffusion of the plasma membrane enzymes 5' nucleotidase, ATP-ase and alkaline phosphatase during incubation. 2. Treatment of fixed sections with different kinds of buffer at 37 degrees C induces diffusion of enzyme activity from the plasma membrane to other sites of the section and into the incubation medium. This buffer influence depends on temperature: at 4 degrees C only a slight diffusion occurs. Addition of phospholipase C, digitonin or taurocholate to the buffer opposes the buffer effect. 3. Pretreatment of frozen cryostat sections with a mixture of equal parts of chloroform and acetone give a good fixation of the plasma membrane enzymes 5'-nucleotidase, ATP-ase, alkaline phosphate and leucyl-beta-naphthylamidase. During this treatment the different kinds of lipids present in the membrane are ex-racted equally. After this fixation buffer treatment does not cause a visible diffusion of enzyme activity in the section. Only a slight diffusion (1 till 7 percent) into the buffer solution takes place. 4. The mentioned treatments open up possibilities to get insight into the membrane anchorage of plasma membrane enzymes.

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