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Purification of brush border membrane vesicles from horse kidney cortex using Percoll
M Boudouard1, J Giudicelli, P Sudaka
1Laboratoire de Biochimie, Faculté de Médecine, Nice, France.
Preparative Biochemistry
|January 1, 1988
Summary
Researchers developed a fast method to isolate horse kidney brush border membrane vesicles. This technique uses Percoll-gradient centrifugation to reduce contamination, ensuring a purer sample for study.
Area of Science:
- Biochemistry
- Cell Biology
- Renal Physiology
Background:
- Brush border membrane vesicles are crucial for studying renal transport.
- Previous methods for vesicle preparation were time-consuming or resulted in contamination.
Purpose of the Study:
- To describe a rapid and efficient method for preparing purified brush border membrane vesicles from horse kidney cortex.
- To validate the purity and characteristics of the prepared vesicles.
Main Methods:
- Utilized self-orienting Percoll-gradient centrifugation for vesicle isolation.
- Employed electron microscopy to assess morphological purity.
- Measured L-alanine uptake to confirm functional integrity of the vesicles.
Main Results:
- A rapid preparation method for horse kidney brush border membrane vesicles was established.
- Percoll-gradient centrifugation effectively minimized microsomal membrane contamination.
- Electron microscopy confirmed the structural integrity of the vesicles.
- Vesicle preparations demonstrated active L-alanine transport.
Conclusions:
- The described method provides a fast and reliable way to obtain purified horse kidney brush border membrane vesicles.
- This preparation is suitable for further investigations into renal transport mechanisms.