Phosphoprotein Enrichment Kit, 635624, Takara Bio Inc (CAT#: STEM-GT-3072-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: 6 Preps

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

MWCO Ultrafiltration Spin Units, 12 mL, Pack Of 24, MWCO 50 K, 17-0291-50, Marvelgent biosciences (CAT#-STEM-GT-3055-LGZ)
MWCO Ultrafiltration Spin Units, 12 mL, Pack Of 24, MWCO 100 K, 17-0291-00, Marvelgent biosciences (CAT#-STEM-GT-3056-LGZ)
Capturem™ His-Tagged Purification Miniprep Kit, 20 Purifications , 635710, Takara Bio Inc (CAT#-STEM-GT-3057-LGZ)
Capturem™ His-Tagged Purification Maxiprep Kit, 6 Purifications , 635713, Takara Bio Inc (CAT#-STEM-GT-3058-LGZ)
Capturem™ His-Tagged Purification Large Volume, 4 Units, 635724, Takara Bio Inc (CAT#-STEM-GT-3059-LGZ)
Capturem™ His-Tagged Purification 24-Well Plate, 1 x 24-Well Plate, 635730, Takara Bio Inc (CAT#-STEM-GT-3060-LGZ)