• method developments to extract proteins from oil palm chromoplast in Tanzania
  • method developments to extract proteins from oil palm chromoplast in Tanzania
  • method developments to extract proteins from oil palm chromoplast in Tanzania
  • method developments to extract proteins from oil palm chromoplast in Tanzania
  • What is the primitive method of plant ECM protein isolation?
  • Perhaps the primitive method of plant ECM protein isolation involves the direct extraction in solution containing CaCl 2 gradients and plasmolysis of plasma membrane to shrink ECM [ 11, 12 ].
  • What is the protocol for ECM extraction & protein extraction?
  • The protocol for ECM isolation, protein extraction , and mass spectrometry coupled gel-based analysis described in this chapter is simple, reproducible, robust, and easy to carry out, that yield intact and pure ECM fraction and ECM proteins.
  • What are the caveats of plant ECM protein extraction & isolation?
  • Extraction and isolation of plant ECM proteins features key caveats due to the lack of adjoining membrane, the presence of a polysaccharide–protein network that traps contaminants, and the existence of high phenolic content.
  • How are ECM proteins extracted?
  • The major steps involved mechanical disruption, density-based gradient centrifugation and series of stringent washes to remove contaminating proteins and extraction of ECM proteins with commonly used wall protein extractants, CaCl 2 (Subheadings 3.1 and 3.3 ). Rigorous purity assessment is performed at organellar and protein levels.
  • How are ionically bound ECM proteins extracted?
  • Ionically bound ECM proteins were eluted by incubating cultured cells in 1 M NaCl buffer followed by homogenization, vacuum infiltration, and extraction. 0.2 M CaCl 2 and potassium phosphates were used to extract tightly ionically bound (fraction 2) ECM proteins.
  • How to separate ECM protein from insoluble fraction?
  • Shake the ECM pellet with extraction buffer on a shaking platform for 2 h at 4 °C to remove any further contaminants and to release ECM proteins ( see Note 16 ). 4. Following centrifugation at10,000 × g for 10 min at 4 °C, the ECM proteins gets separated from the insoluble fraction ( see Note 17 ).