Plant cells contain several types of intracellular membrane-bound structures, known as organelles, such as nucleus, mitochondrion, lysosome, endoplasmic reticulum, Golgi apparatus, chloroplast, contractile vacuole, peroxisome, ribosome, and so on. Each organelle has its specific function (Table1). Isolation of subcellular organelles from plant cells is prerequisite for various plant biological topics, including functional and structural genomics, as well as proteomic profiling. Lifeasible, as a leading plant biotechnology company in the world, provides one-stop services for separation of different types of plant organelles. Our sample-to-report services cover every key step from sample homogenization and purification, to organelle enrichment and integrity detection.
Table 1. The specific functions and some marker enzymes for major organelles.
|Nucleus||DNA storage; control of all activities of the cell; RNA transcription||RNA polymerase II|
|Mitochondrion||Energy production||Succinate dehydrogenase; cytochrome c oxidase|
|Smooth endoplasmic reticulum||Lipid production; detoxification|
|Rough endoplasmic reticulum||Protein production; in particular for export out of the cell||Glucose-6-phosphatase|
|Golgi apparatus||Protein modification and export|
|Peroxisome||Lipid Destruction; containing oxidative enzymes|
|Lysosome||Protein destruction||Acid phosphatase|
|chloroplast||Photosynthesis; trapping energy from sunlight||Sialyl transferase|
|Vacuoles||Storage; transportation; homeostasis balance||Alkaline phosphodiesterase|
Cell Disruption and Homogenization
The first step of cell disruption is the removal of cell wall by enzymatic digestion and then homogenization of the cells. The homogenization process involves 5 steps: Dounce homogenization, filtration, grinding, sonication, and solubilization. In order to ensure the integrity of organelles, isotonic sucrose is usually added to the homogenization buffer to prevent osmotic rupture of organelle membranes during the homogenization process.
Plant Organelle Purification
Following homogenization, the next step is purifying organelles from homogenates. We provide several methods for isolating subcellular organelles, which include, but not limited to:
Purity, enrichment, and integrity detection of organelles
Evaluation of the integrity, enrichment, as well as the presence of contaminating organelles for target organelles are important for the studying of their various biological processes. Our experts have developed a large variety of methods for analyzing the integrity and purity of plant organelles. The most commonly used methods are microscopic techniques and marker enzymes activity analysis.
With cutting-edge equipment and experts in plant cell biology, we provide you with customized protocols for each specific plant organelle isolation and analysis. Working closely with Lifeasible, you will benefit from our fast, reliable, and cost-effective services for the separation of plant organelles. Welcome to contact us for technical consults and price quotes.