Phosphoprotein Kit - Buffer A, 635626, Takara Bio Inc (CAT#: STEM-GT-3071-LGZ)

The Phosphoprotein Enrichment Kit provides an efficient affinity-based procedure to isolate phosphorylated proteins from mammalian cells and tissues. Each kit includes a complete set of buffers, and 6 high-capacity columns for the enrichment of cytoplasmic and membrane-bound phosphoproteins regardless of modified amino acids, including serine, tyrosine, or threonine.

Our enrichment program offers many advantages:

The process is fast; the average cell-to-sample purification time is less than 2 hours.
It is also simple and consists of four main steps: adding extraction/loading buffer to the cell or tissue pellet to extract total cellular protein, loading the extract onto an affinity column, washing, and finally washing with detergent-free The elution buffer elutes the bound phosphorylated protein.
A separate buffer - the extraction/loading buffer - is used for the protein extraction and affinity column steps, making buffer exchange unnecessary. This saves time and prevents sample loss.
The process is non-denaturing, so phosphoproteins remain folded throughout the process, even during extraction and elution steps.
Each column has a maximum binding capacity of ~4 mg of phosphorylated protein.

Highly selective enrichment of phosphoproteins

The Phosphoprotein Enrichment Kit can be used with any mammalian cell type. Cell lines tested include NIH 3T3, HEK 293, HeLa, Cos-7 and Jurkat. The enrichment process was highly efficient, as demonstrated by Western blotting analysis. Using a colorimetric phosphate detection method, we found the majority of phosphoproteins in the eluate; negligible traces were detected in the wash fractions.

Phosphoprotein affinity columns produce concentrated phosphoprotein solutions that can be analyzed by several different methods, including mass spectrometry and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE).

Size: 500 mL

Non-denaturing protocols preserve protein conformation and solubility
Ideal for many downstream applications including mass spectrometry and 2D-PAGE
Improving the Sensitivity of Phosphorus Detection Experiments

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