Protein Extraction Efficiency of Soft and Hard Seeds using the Precellys Lysing Kits - Bertin Instruments

    Agronomy

    Protein Extraction Efficiency of Soft and Hard Seeds using the Precellys Lysing Kits

    Sources: Department of Chemical Physiology, The Scripps Research Institute, La Jolla, CA

    Context

    Sample preparation for protein extraction is a critical first step to achieve reliable analytical results. In this study we compared different bead kits using two different suspension liquids and two kinds of plant organ, soft and hard seeds, namely peanut and rice, respectively.

     

    Results

    Table 1

    Table 1. Protein extraction efficiency as a percentage of total protein content, using saline extraction buffer. Extraction efficiency is based on the theoretical amount of protein, 25.8g, 6.12g/100g of peanut, rice respectively. (USDA National Nutrient Database for Standard Reference Release 26, Feb 7, 2014. http://ndb.nal.usda.gov/ndb/foods). **A MudPIT method for LC-MS/MS was used to identify proteins from rice (Delahunty CM, Yates JR. MudPIT: multidimensional protein identification technology. Biotechniques 2007; 43: 563-569).

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    Figure  1.    Protein  concentration  was  quantified  in  the medium  aqueous  phase  after  peanut  (A)  and  rice  (B) homogenization in saline buffer (green bars) versus ddH2O (blue bars) using 4 different lysing matrices.

    Protein  extraction  efficiency  in  peanuts  and  rice  was compared  after  bead-beating  with  4  different  lysing kits.  The  use  of  saline  buffer  increased  protein extraction  efficiency  as  expected  (Robert  LS, Nozzolillo  C,  Altosaar  I.  Homology  between  legumin-like  polypeptides  from  cereals  and  pea.  Biochem  J 1985; 226: 847-852).  The CK14 beads gave the best protein yields compared to the other lysing beads for both hard and soft seeds.

     

     

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