PhosPro - Phosphoprotein enrichment kit 10rxns

In stock
SKU
P5012-10
375,00 €
PhosPro - Phosphoprotein enrichment kit 10rxns
£340 / €375
Supplier
Genomine
Cat. no.
P5012-10
Kit size
10
Links
Info & Protocol

Features

  • Phosphoprotein specific: Highly selective enrichment of phosphoprotein from complex mixtures of cell lysate
  • Sensitive
  • Convenient
  • Reproducible
  • Enrichment from native or denatured protein solution.

Protein phosphorylation is one of the most frequently occurred posttranslational modifications and plays a critical role in cellular regulatory events. Most cellular processes are in fact regulated by the reversible phosphorylation of proteins on serine, threonine and tyrosine residues. In fact, phosphorylation of proteins plays a key role in oncogenesis, cell signaling, apoptosis and immune disorders1. Despite the importance and widespread occurrence of this modification, profiling of phosphoproteins in cells is still a challenge, due to the low copy of phosphorylated proteins in cell and the relative amount of phosphoproteins compared to unphosphorylated proteins.

Radiolabeling by 32P labeling is frequently used conventional method for investigation of phosphoprotein profile in conjunction with 2-DE or 1-D gel electrophoresis and autoradiogram. Alternatively, western blot analysis probed by phosphoprotein-specific antibody is also used for this purpose. Mass spectrometry has been shown to be a reliable and routine tool to identify proteins in a high throughput manner. However, the identification of phosphorylation by mass spectrometry is not a trivial matter and to this day is not routine also due to the low copy of phosphorylated proteins in cells.

This phosphoprotein enrichment and exclusion of unphosphorylated proteins provides advanced chance in detecting protein phosphorylation in gels with non-radiolabeling method(eg. Staining with fluorescence dye) and enables quantitative comparison between cells.
Procedure Summary:

1. Native & denatured Protein extraction

2. Phosphoprotein specific complex forming

3. Precipitation of phosphoprotein complex

4. Dedelipidation and recovering of phosphoprotein

Average Yields Specificity: 100% Selectivity: >87% Sensitivity: >93% / 50ng Yield: >93%
Lysis buffer 10 ml
Dilution buffer 20 ml
Solution A 3 ml
Solution B 5 ml
Disolving solution 8 ml
Delipidation solution 10 ml
2-DE analysis of enriched phosphoprotein fraction from total denatured(A,C) & native protein(B,D) stained with CBB(A,B), and stained with ProQ Diamond staining(C,D).


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