Specifications

Applications- SDS-PAGE, Mass Spectrometry, etc.

Sample Type- Formalin Fixed Paraffin Embedded tissue

Sample Size- five ̴10u thick sections

Yield- up to 10,000ug/mL of protein

Kit Sizes- 10 Assays, 20 Assays

Product Details

ITSI’s Total Protein Isolation Kit for Formalin Fixed Tissue Kit ToPI-F2 contains in a ready to use format all the reagents necessary to extract protein from 10 or 20 specimens, (each consisting of five ̴10u thick sections each) depending on the kit sized selected and our validated procedure which will guide you through the process. From each specimen of 5 sections up to 10,000 ug/ml of protein can be extracted, dependent on the type of tissue, length of storage and fixation technique used.

Principle

Developed specifically to extract proteins from Formalin Fixed Paraffin Embedded (FFPE) samples for use in 1D PAGE and mass spectrometry applications ToPI-F2 has been used to extract proteins from samples stored for up to 10 years. ToPI-F2 allows valuable proteomics data to be derived from one of the most extensive archives of pathology samples in existence, one that continues to grow massively every year.

Procedure

Samples are deparaffinized in a series of solutions over several incubations. After the final incubation and centrifugation the extracted proteins are present in the supernatant ready for the next step of analysis.

Applications

Expanding from histochemical analysis of FFPE specimens into proteomic analysis of is highly desirable because FFPE Specimens make up the largest and longest time-spanning collections of patient materials in pathology archives around the world. With the addition of the clinical records that are available, and the clinical behavior and outcomes for many of the cases, FFPE specimens represent a wealth of data for translational cancer studies, biomarker identification, and comparative proteomics.

Supporting Data and Figures

Fig 1: 1D-PAGE gel image of ToPI-F2 Human Lung Tissue1D-PAGE gel image of protein isolated from formalin-fixed paraffin embedded (FFPE) human lung tissue using the ToPI-F2 kit. MW is molecular weight marker and lanes 1-4 are proteins isolated from four different lung specimens. Approximately 8ug of total protein were loaded per lane.

Each kit contains optimized and ready to use reagents including:

• Solution 1
• Solution 2
• Solution 3
• Microfuge tubes
• Validated protocol