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manufacturer: Nard Institute manufacturer: MANAC Incorporated

New Method for Analysis of Phosphorylation Molecules
Phos-tag™ series

Phos-tag™ is a functional molecule that binds specifically phosphorylated ions.
It is applicable for the specific separation of phosphorylated proteins (Phos-tag™ Acrylamide) as well as for the detection using western blot (Phos-tag™ Biotin), enrichment, separation and purification (Phos-tag™ Agarose), and MALDI-TOF/MS (Phos-tag™ Mass Analytical Kit).

Methods for determining the phosphorylation status of proteins (i.e., phosphoproteomics) are thus very important with respect to the evaluation of diverse biological and pathological processes.
In 2002, Prof. Koike's group (Hiroshima University) reported that a dinuclear metal complex (i.e., 1,3-bis[bis(pyridin-2-ylmethyl) amino]propan-2-olato dizinc(II) complex) acts as a selective phosphate-binding tag molecule, Phos-tag™ in an aqueous solution at a neutral pH (e.g., Kd = 25 nM for phenyl phosphate dianion, Ph-OPO32-).

Basic structure of Phos-tag™

basic structure of Phos-tag
      M2+: Zinc ion or Manganese ion

LINK to Phos-tag PAGE applications
Phos-tag was advertised in nature
on Sep. 25, 2104 and Jan. 29, 2015.
⇒ zoom

1. Features

  • Selectivity of binding of a phosphate ion(2-) is much higher than that of other anions.
  • A stable complex is formed under physiological conditions (pH 5 to 8).
  • Recognition of all phosphorylated forms of Ser / Thr / Tyr
  • Radioisotope labeling is not required.
  • Downstream applications followed by mass analysis, etc. are applicable.

2. Phos-tag Series List

♣ 1. Phos-tag™ Acrylamide

Providing a phosphate affinity SDS-PAGE for mobility shift detection of phosphorylated protein.

      LINK to
Phos-tag™ Acrylamide

♣ 2. SuperSep Phos-tag™

Ready-to-Use Precast Phos-tag™ Acrylamide Gels are available!

      LINK to
SuperSep Phos-tag™

♣ 3. Phos-tag™ Biotin

  • Sensitive Detection of Phosphorylated Proteins on PVDF membrane
  • Downstream procedure such Sensitive Detection of Phosphorylated Proteins on PVDF membrane
      LINK to

      New Product: A higher sensitive type Phos-tag™ Biotin BTL-111   1 mM Aqueous Solution

Phos-tag™ Biotin

♣ 4. Phos-tag™ Mass Analytical Kit

  • Suitable for mass analysis.
  • An equivalent amount of acetate anion binds to Phos-tag™ MS-101
  • Anion-free Phos-tag MS-101 is a 3+ ion in aqueous solution.
      LINK to
Phos-tag™ Mass Analytical Kit

♣ 5. Phos-tag™ Agarose

(AMP2- binding site = 3 - 5 μmol/mL-gel)

  • providing an efficient procedure for separation and concentration of native phosphoproteins and phosphorylatd peptides from biological sample at physiological pH.
  • It corresponds to a variety of column scales and applications depending on the amount and the purpose.
      LINK to
  • Manual:   ⇒ Enrichment of Phosphorylated Proteins from Cell Lysate
    -Phosphate Affinity Chromatography using Phos-tag™ Agarose
    (version 8, May, 2015) PDF (1.8 MB/11 p)
  • Leaflet updated in September, 2016 ⇒ PDF PDF (1,113 KB/4 pages)
Phos-tag™ Agarose

3. References

References on Phos-tag™ Chemistry
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