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Phos binding reagent acrylamide
An equivalent to Phos-tag acrylamide. Separation of phosphorylated and non-phosphorylated proteins

Catalog No.F4002
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Sample solution is provided at 25 µL, 10mM.


Protein phosphorylation is an important covalent post-translational modification that can alter the structural conformation of a protein, which then regulates the function, location and specific binding of the target protein. Many cellular processes are regulated by the reversible phosphorylation of proteins and 30% of the proteins are likely to be phosphorylated at some point during their existence.

The determination of the phosphorylation state of proteins is important for defining protein kinase substrates and revealing the activation state of signal transduction pathways. Methods for determining the phosphorylation status of proteins are thus have important implications in the understanding of diverse biological and pathophysiological processes, such as signal transduction pathways, cancer and other diseases.

Phos binding reagents are specific and selective phosphate-binding reagents, and exhibit preferential ionic interactions with phosphorylated ions on phosphorylated proteins or peptides at neutral pH. Phos binding reagent is a dinuclear metal (Zn2+ or Mg2+) complex.

Phos binding reagent acrylamide (PBR-A) is a specific reagent for separation of phosphorylated proteins using SDS-PAGE method.

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Basic structure of Phos binding reagents

• <Zn2+-Phos binding reagents>: a stable complex is formed with a phosphate group at the ratio of 1 to 1 at pH 5 ~ 8.

• <Mn2+-Phos binding reagents>: a stable complex is formed with a phosphate group at the ratio of 1 to 1 at around the pH 9.


1. Recognition of all phosphorylated forms of Tyr/Ser/Thr.

2. Followed by Western blotting and Mass analysis.

3. Simultaneous detection of phosphorylated/non-phosphorylated proteins using the total antibody without the anti phosphorylated protein.

4. Simply add Phos binding reagent acrylamide and MnCl2 solution* to acrylamide solution in the preparation of SDS-PAGE gel.

(*: Add 5.0 mM Phos binding reagent Acrylamide Aqueous Solution and 10 mM MnCl2 solution in equal amount in the gel).


1. M. J. Hubbard, P. Cohen. On target with a new mechanism for the regulation of protein phosphorylation. Trends Biochem. Sci. 18: 172 (1993).

2. B. Agnew et al. Compositions and methods for detection and isolation of phosphorylated molecules. US Patent # 7,102,005. (September 5, 2006).