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Plant Proteomics

CelLytic P

Description: This is a non-ionic detergent-based reagent, which offers a convenient method for efficient plant cell lysis and protein solubilization. It's a non-denaturing reagent and maintains protein immunoreactivity and biological activity (Fig. 1). CelLytic P is efficient, rapid, and ready to use. It contains bicine buffer, which is preferable for many biological activities. Use of CelLytic P enables extraction of proteins from less than one gram to hundreds of grams of fresh or frozen leaves, employing the same short procedure. It has been tested on (but not limited to) four plant models: tobacco, tomato, spinach, and arabidopsis.
 
The resulting lysate is suitable for:
  • Western blot analysis
  • Protein staining such as Coomassie and silver staining
  • Enzymatic assays: e.g. Kinase assays (tested for tyrosine and serine kinase activity) and phophatase assays (tested for general and alkaline phosphatase)
  • DNA:protein interaction assays
    (Gel-shift)
Detection of DNA/Protein interactions
Figure 1. Compatibility with Gel Shift Assay.
Protein extracts were prepared with the CelLytic-P reagent from spinach leaves. A double stranded 32P labeled CREB oligonucleotide probe was incubated with 28 µg of the whole cell extract (lanes 2-5) or without whole cell extract (lane 1, free probe). Binding reactions with the extracts were performed in the absence [-] of competitor oligonucleotide (lanes 1-2) or in the presence of x100 or x500 fold excess of unlabeled CREB binding motif oligonucleotide (specific competitor [SP], lane 3 and lane 4, respectively) or in the presence of x100 fold excess of unlabeled oligonucleotide (non specific competitor, [NS] lane 5). Binding reactions were run on a non-denaturing 6% polyacrylamide gel, dried and imaged on X-ray film. The arrows indicate the CREB-DNA complex and the free probe.

Ordering Information

Product Product Name Package Size Technical Bulletin
C2360 CelLytic P Cell Lysis Reagent 50 mL
250 mL
X