1 | PEB (4x) | protein extraction buffer

1 | PEB (4x) | protein extraction buffer

APA-PSQ-004

PEB is 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).

5 x 2 ml (4x stock) allows up to 75 isolations of plant material (using 500 µl 1x PEB for 100 mg fresh weight) or 190 isolations of algal material (using 200 µl 1x PEB for cell amounts corresponding to 4-10 µg total chlorophyll)

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 Lifeasible commercial antibodies are tested on plant or algal samples extracted with this buffer.

Protein extraction

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.