Extraction, Purification and Analysis of Plant Endoplasmic Reticulum

Endoplasmic reticulum (ER) is a continuous membrane system composed of smooth endoplasmic reticulum (sER) and rough endoplasmic reticulum (rER) attached with ribosomes. Endoplasmic reticulum membrane accounts for about 50% of the total intracellular membrane area, which is the largest proportion of the membrane structure in eukaryotic cells. ER is involved in protein synthesis, folding, and stress response under adverse conditions. The study of endoplasmic reticulum proteomics is the basis for revealing the mechanism of subcellular stress resistance. Prior to mass spectrometry, the purity of subcellular fractions should be evaluated to confidently conclude the presence of relevant specific protein forms and deepen the understanding of their biological functions.

Lifeasible, as a leading global company, is committed to helping our customers achieve effective and successful research. We provide extraction of endoplasmic reticulum, purity assessment, and proteomic analysis in plants. In addition, we deliver reliable results and reports on time to our customers worldwide.

Extraction of Endoplasmic Reticulum in Plants

• To further explore the function of the plant endoplasmic reticulum, it is necessary to isolate endoplasmic reticulum from plant tissues. Currently, we provide ER isolation and purification services for plant root tips, seeds, internodes, leaves, and other tissues.
• We have developed a number of methods to enrich ER components, such as continuous iododiol gradient, discontinuous sucrose gradient, continuous sucrose gradient, and ultracentrifugation combined or without using sucrose gradient. Although these methods are feasible, the process is quite complicated and time-consuming. Therefore, we also developed ER enrichment kit for the efficient enrichment of plant ER.

Fig.1 Extraction and protein purification of endoplasmic reticulum in plants.

Purity Assessment of Plant Endoplasmic Reticulum

• We provide purity assessment of ER in plants by microscopy, immunoblotting, and enzymatic analysis.
• The integrity and purity of the target fractions were determined by electron microscopy. However, it relies heavily on commercial dyes developed for specific subcellular organelles. To overcome this shortage, proteomic analysis of ER, immunoblotting, and enzymatic analysis were used as alternative methods to assess ER purity.
• Antibodies for immunoblot analysis and manufactured proteins for enzymatic analysis have been reviewed to indicate contamination from the cytoplasm, plasma membrane, nucleus, mitochondria, and chloroplasts.

Proteomic Analysis of Plants Endoplasmic Reticulum

• Protein function depends not only on its molecular structure but also on its subcellular localization and post-translational modification (PTM). Liquid chromatography-mass spectrometry (LC-MS) has become the method of choice for spatial proteomics (localization of proteins and their dynamics at the subcellular level) and PTMs due to its unique measurement capabilities.
• We provide plant ER proteomic analyses that help unravel cellular signaling networks and protein-protein interactions and provide a molecular understanding of the mechanisms involved in biological stress responses.

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