Application

The plant total Protein Extraction Kit has been used to isolate crude seed protein. It has also been used to extract the total protein.

General description

The Plant Total Protein Extraction Kit is designed specifically for use in plant bioscience to extract a qualitative sample of all proteins from any type of plant species or tissue. The protocol does not require any ultracentrifugation or aqueous polymer two-phase partitioning (APTP). The kit includes two reagents; a plant specific protease inhibitor cocktail and a new chaotropic reagent with increased solubilizing power to extract more hydrophobic proteins (C0356, Protein Extraction Reagent Type 4). Routine use of the reagent in sequential protein extraction of a plant tissue sample with fresh reagent removes nearly all protein from the tissue. The protease inhibitor cocktail is a mixture of protease inhibitors with broad specificity for the inhibition of serine, cysteine, metalloproteases, aspartic, and aminopeptidases. It contains pepstatin A, 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), E-64, leupeptin, 1,10-phenanthroline, and bestatin. The cocktail has been demonstrated to be highly effective in preventing protein degradation during the extraction process. Following removal of polyphenolics, tannins, and other interfering substances, ground plant tissue, fresh or frozen, is resuspended in the chaotropic reagent. Plant debris is pelleted by centrifugation and the protein extract is collected. The end result is a qualitative total protein sample, ready for downstream proteomic analysis.

Quantity

Sufficient for preparation of 20 separate extractions from plant tissue samples of 10-250mg