1 | PEB (4x) | protein extraction buffer

1 | PEB (4x) | protein extraction buffer

• Background: PEBis an extraction buffer for disruption and solubilisation of total protein from plant tissue and algal cells. The use of the anionic detergent LDS together with the recommended procedure (combination of sonication and freeze/thaw cycles) has been shown to increase the number of solubilised and non-degraded proteins when compared to other methods of cell disruption (see reference). The estimated hands-on time for the recommended procedure is 20-30 minutes for 1-2 samples. Expected yields will be 1.5-6 µg/µl total protein (recovered from standard procedure) depending on the starting material, e.g. its biological stage, homogenization method used (bead beater vs. sonication).
• Reactivity: PEB has been tested on a wide range of species and tissues from Higher plants, Mosses, Llichens, Algae, Diatoms, Dinoflagellates, Cyanobacteria. Extracts may be quantified using detergent (LDS) compatible methods, and have been shown to give highly reproducible and quantitative results in subsequent SDS PAGE gel electrophoresis, Western blotting, and Immunoprecipitation (IP).Most of Agrisera commercial antibodies are tested on plant or algal samples extracted with this buffer. An example can be foundhere.
• Applications: Protein extraction
• References & Citations: Huanget al. (2023) Along with cyclic electron flow and non-photochemical quenching, water-water cycle is involved uniquely in alleviating Zn stress-caused photodamage in Melia azedarach. Tree Physiol. 2023 Apr 18:tpad045. doi: 10.1093/treephys/tpad045.Altuntaset al. (2020). Proline-stimulated signaling primarily targets the chlorophyll degradation pathway and photosynthesis associated processes to cope with short-term water deficit in maize. Photosynth Res. 2020 Apr;144(1):35-48. doi: 10.1007/s11120-020-00727-w.Pérez-Lópezet al. (2020). Transcriptome Analysis Identifies Plasmodiophora brassicae Secondary Infection Effector Candidates. J Eukaryot Microbiol. 2020 Jan 11. doi: 10.1111/jeu.12784.Morinet al. (2019). Morin et al. (2019). Response of the sea-ice diatom Fragilariopsis cylindrus to simulated polar night darkness and return to light. Limnology and Oceanography. 9999, 2019, 1â??20. (sea-ice diatom)Bausch,A.R., Juhl, A.R., Donaher, N.A. et al. Mar Biol (2019) 166: 80.Matsuoand Atsumi (2018). Xylosylation of proteins by expression of human xylosyltransferase 2 in plants. J Biosci Bioeng. 2018 Sep;126(3):371-378. doi: 10.1016/j.jbiosc.2018.03.013.Brouweret al. (2011) TheImpact ofLightIntensity onShade-InducedLeaf Senescence. Plant Cell Environ. Dec. 15 (ahead of print).Kosawanget al. (2011) Hydrogen yield from a hydrogenase in Frankia R43 at different levels of the carbon source propionate. Journal of Environmental Management, Jan 26
• Storage Conditions: Stable at RT for at least 1 month; short-term storage (6 monthss) at 4°C and long term storage (1 year or more) at -20°C.