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Used together with the Stabilizor system, the Stabilizor 2D-GE Extraction Kit (Tissue) maximizes the recovery of proteins from heat-stabilized tissue to obtain a representative picture of protein states at the moment of sampling. The kit contains a validated protocol, accessories and solutions needed to homogenize and extract proteins prior to 2D-GE.
Produce high quality samples for consistent 2D-GE analysis
Reduce time to results
Follow a ready to use, validated protocol - no need for optimization
Use standard laboratory equipment
The method used to extract protein components from the heat-stabilized tissue before analysis must ensure effective re-solubilization of protein components and disaggregation of any protein-protein complexes that may have formed.
The extraction protocol and buffer supplied in the Stabilizor 2D-GE Extraction Kit were optimized and validated by comparison with commonly-used protocols and buffers. Results from protein concentration measurements, 1D and 2D-GE analysis confirmed that ultra-sonification during homogenization, in buffer containing thiourea, CHAPS and DTT, improved re-solubilization of proteins.
2D-GE analysis of heat-stabilized rat liver prepared using the Stabilizor 2D-GE Extraction Kit shows good recovery of high and low molecular weight molecules and demonstrates efficient extraction from a sample known to contain high levels of basic proteins. Gel evaluation and analysis performed on Progenesis SameSpots version 4.5, Nonlinear Dynamics.
During protocol development a ratio of 10:1 buffer:sample, i.e. 10 μl buffer to each mg tissue, gave the most effective extraction and achieved highest protein concentration. Normal or heat-stabilized tissue (chicken liver) samples were processed identically using only different buffer:sample ratios.
Results from a recent publication (Biomarkers of disease and post-mortem changes — Heat stabilization, a necessary tool for measurement of protein regulation, Kultima K, et al, J Prot, 2011) demonstrates a remarkable overlap between proteins commonly found expressed differentially in disease states in 2D-GE proteomics studies and proteins found to change post-mortem due to sample handling.