NoviPure Soil Protein Kit

NoviPure Soil Protein Kit
Extract protein from all soil types.
Size/Component Catalog #
20 preps 30000-20
Product Description

Details

  • Cleaner Protein - Reduces humic substance interference
  • Higher Yields - Efficient of intracellular and extracellular microbial protein from only 5 grams of soil
  • Accurate results - In mass spectrometry, 1D and 2D gel electrophoresis and other proteomics applications 

The NoviPure Soil Protein Kit is designed to extract extracellular and intracellular microbial protein from a wide range of soil types without co-extraction of interfering compounds such as humic substances. The patent-pending, two buffer protocol utilizes bead beating with a mixture of glass and ceramic beads to efficiently lyse cells while solubilizing intacellular as well as extracellular protein. The end result is a protein pellet with considerably fewer impurities when compared to traditional methods. The protein can be resuspended in any buffer desired for further anlysis or storage. Protein extracted with the NoviPure Soil Protein Kit has been used successfully in 1D and 2D polyacrylamide gel electrophoresis and mass spectrometry (2D LC-MS/MS). All reagents and plastics are certified protease- and protein-free to protect valuable samples. This kit is optimized for the of denatured protein from soil microbes.

Specs

Specs

Format Patent-pending two buffer extraction method
Method Bead Beating
Sample Size 2 - 5 grams
Throughput 1-2 samples
Storage Store at room temperature (15-30°C)
Pre-chill Solution SP1 and Solution SP2 before use.
Bead Type 0.1 mm glass, 0.1 mm ceramic mix
Sample Types Processed All types of soils, sand, and sediment
Equipment Required Vortex and Vortex Adapter (13000-V1-50)
Refrigerated Centrifuge for 50 ml tubes
Refrigerated Centrifuge for 2 ml tubes
Catalog # 30000-20, 30000-S
Old Name NoviPure Soil Protein Extraction Kit
SDS/Protocol

SDS/Protocol

Protocol

SDS 

Data

Data

Cleaner Protein
 
    Figure 1. Cleaner protein is observed in compost samples extracted with NoviPure Soil Protein Kit. (A) Total protein was extracted from 5 g of sterile compost samples spiked witth C. albicans using either the NoviPure Soil Protein Kit (samples 1-2) or a method employing sucrose and phenol extractions1 (4, 5). A pure C. albicans culture was extracted with the NoviPure Soil Protein Kit (3) or sucrose/phenol as a control (6). (B) Total protein was extracted from 5 g sterile compost samples spiked with E. coli using either the NoviPure Soil Protein Kit (samples 1-2) or a method employing SDS and boiling2 (3, 4). Proteins (20 µl per sample) were run on a 1D SDS-PAGE gel and visualized using stain-free gels3. In each case, samples extracted with the NoviPure Soil Protein Kit contained fewer humic substances as evidenced by reduced background smearing, making it possible to visualize protein bands on a 1D gel.
 
Higher Protein Yields
 
  Figure 2. High protein yields are observed from challenging soil types using the NoviPure Soil Protein Kit. Total protein was extracted from 5 g samples of sterile soil and sediments spiked with E. coli using the NoviPure Soil Protein Kit. Proteins (20 µl per sample) were run on a 1D SDS-PAGE gel and visualized using stain-free gels3. Protein bands were easily visualized in all cases, and matched the E. coli culture control. (1, 2) - Freshwater lake sediment; (3, 4) - Lagoon sediment; (5, 6) - Beach sand; (7, 8) - Agricultural soil; (9, 10) - Pure E. coli culture.
  
Successful Results 
  
  Figure 3. Successful results in 2D Gel Electrophoresis using protein extracted with the NoviPure Soil Protein Kit. Protein was extracted from 5 g sterile compost samples spiked with E. coli using the NoviPure Soil Protein Kit. Protein (20 ul per sample), was analyzed by 2D SDS-PAGE in the pH range 3-10 and visualized using silver staining. Protein was easily visualized with no interference from humic substances.
 
Accurate Results
 
  Figure 4. Accurate results in 2D LC - MS/MS using protein extracted with the NoviPure Soil Protein Kit. Total ion chromatogram data shing base peaks of protein. Image is fraction 4 of SCX cation exchange. Protein was extracted from 5 g sterile compost samples spiked with E. coli using the NoviPure Soil Protein Kit. Proteins were digested adn peptide enriched using reverse phase ion exchange and analyzed with 2D LC - MS/MS. Raw spectra were searched against SwissProt protein database (March 2013 release). 1076 unique proteins were identified.
 

1Taylor & Williams, 2013 Microb. Ecol. 59:390-99

2Chourey et al. 2010 J. Proteome Res. 9:6615-22

3BIORAD mini PROTEAN® TGX Stain-Free™ Gels