Phos-tag™ Fluorescent Gel Stain Reagents

• Highly sensitivity
• Staining at physiological pH
• High selectively to pSer, pTyr, pHis, and pAsp residues
• No need for any radioactive material, labeling with a chemical or antibody
• Completed within 2 hours

• Competitor Product A

  • pH：2-4
  • 5 steps
    (fixing・washing・staining・destaining・washing)
  • The time required：≧ 5 hours
  • Solution exchange：11 times
  • The detection limit of ovalbumin：>5 ng/lane

• Phos-tag™ Fluorescent Gel Stain Reagent

  • pH：7-8 (no dephosphorylation occurs)
  • 3 steps
    (fixing・staining・washing)
  • The time required：≦ 2 hours
  • Solution exchange：4 times
  • The detection limit of ovalbumin：>1 ng/lane

• Configuration of Equiliprating & Washing Solution as well as Staing Solution requires the Mixed reagents for Phos-tag™ Common Solution 5x (product code：383-15231).

“Quantitative monitoring of His and Asp phosphorylation in a bacterial signaling system by using Phos-tag Magenta/Cyan fluorescent dyes”, Emiko Kinoshita-Kikuta, Hiroshi Kusamoto, Syogo Ono, Keisuke Akayama, Yoko Eguchi, Masayuki Igarashi, Toshihide Okajima, Ryutaro Utsumi, Eiji Kinoshita, Tohru Koike, Electrophoresis, 2019

Staining phosphorylated and non-phosphorylated proteins

• 

• Lane No. ( ) indicates the number of Phosphate groups

  1. Marker
  2. α-casein (9P)
  3. ALP-treated α-casein
  4. β-casein (5P)
  5. ALP-treated β-casein
  6. Ovalbumin (2P)
  7. ALP-treated ovalbumin
  8. Pepsin (1P)
  9. ALP-treated pepsin
  10. ß-galactosidase
  11. Bovine serum albumin
  12. Carbonic anhydrase

Selectivity of the Phos-tag™ Yellow stain: A 10% (w/v) polyacrylamide gel containing four sets of intact and ALP(alkaline phosphatase)-treated phosphoproteins (1 μg each of α-casein, β-casein, ovalbumin, and pepsin) was stained with Phos-tag™ Yellow (left) and subsequently with CBB G-250 gel satin (right).

Data provided by Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike, Department of Functional Molecular Science, Institute of Biomedical and Health Sciences, Hiroshima University

Analysis of phosphorylation of histidine and aspartic acid residues by Phos-tag™ fluorescent gel staining

Signal transduction from EnvZ, histidine kinase, to OmpR: The regulator responses play a role in bacterial gene expression. Phosphorylation of histidine and aspartic acid residues were analyzed on autophosphorylated EnvZ and OmpR phosphorylated by EnvZ.

Data provided by Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike, Department of Functional Molecular Science, Institute of Biomedical and Health Sciences, Hiroshima University

Screening for histidine kinase inhibitors

Screening for histidine kinase inhibitors was performed by using Phos-tag™ fluorescent gel staining.

Data provided by Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike, Department of Functional Molecular Science, Institute of Biomedical and Health Sciences, Hiroshima University