Phos-tag™ PAGE GUIDEBOOK

Phos-tag™ Series

Product Name Pkg. Size Wako Cat. No. Applications
Phos-tag™ Biotin BTL-104 10 mg 301-93531 Specific detection without any anti-phosphorylated antibodies on Western blot. Detection is possible regardless of type of phosphorylated amino acid.
Phos-ta™ Biotin BTL-105 10 mg 308-93541
Phos-tag™ Biotin BTL-111
1mM Aqueous Solution Ref.1)
0.1 mL 308-97201 BTL-111 offers higher sensitivity than BTL-104.
Phos-tag™ Mass Analytical Kit 1 Kit 305-93551 Analysis by MALDI-TOF/Mass
Phos-tag™ Agarose 0.5 mL 302-93561 Enrichment, separation and purification of phorphorylated proteins using column chromatography
3 mL 308-93563

Phos-tag™ Biotin −Detection of phosphoprotein for Western blotting−

This is biotin-bound Phos-tag™ used for detection of phosphoprotein by Western blotting.

Features

● All phosphoprotein can be detected.

● Procedures of experiment are similar to those in ordinary Western blotting.

It can be conveniently used even when target anti-phosphorylated Thr/Ser antibody is not available !

※BTL-104, BTL-105, and BTL-111 have linkers with different lengths. Although the usage of BTL-104 and BTL-105 are similar, BTL-104 is recommendable as the first choice because of its high solubility. BTL-111 offers higher sensitivity.

*Reference of BTL-111

1) Highly sensitive detection of protein phosphorylation by using improved Phos-tag Biotin, Proteomics, 12(7), 932-7 (2012), Kinoshita E, Kinoshita-Kikuta E, Sugiyama Y, Ozeki T, Koike T.

Schematic of western blotting using Phos-tag™ Biotin

Schematic of western blotting using Phos-tag™ Biotin

Phos-tag™ Mass Analytical Kit −Detection Sensitivity of MALDI-TOF/Mass is improved−

Before use, Phos-tag™ Mass Analytical Kit is mixed with samples for MALDI-TOF/Mass analysis. Phosphorylated molecule-Phos-tag™ complex is detected in a positive mode, and hosphorylated molecule usually difficult to detect can be detected with improved sensitivity.

Features

● Detection sensitivity of phosphorylated molecule is improved.

【Kit Contents】
Phos-tag™ MS-101L ……5mℊ
 ([C27H29N6O64Zn2]3+ MW:581.4)
Phos-tag™ MS-101H ……5mℊ
 ([C27H29N6O68Zn2]3+ MW:589.4)
Phos-tag™ MS-101N ……10mℊ
 ([C27H29N6OZn2]3+ MW:584.3)

Phos-tag™ Agarose −Purification of phosphorylated proteins by affinity chromatography−

Fill Phos-tag™ Agarose in a column for use. It can be used for separation, purification and concentration of phosphorylated proteins. Because it is free from surfactants or reducing agents, phosphorylated protein can be obtained in a condition similar to in vivo one.

Features

● Phosphorylated proteins can be purified in 1 hour.

● The proteins can be trapped in physiological condition (pH 7.5).

● Purified with no reducing agent or surfactant.

Phos-tag™ Agarose

  • Lysate was applied on a column filled with Phos-tag™ Agarose. Phosphorylated proteins were concentrated in absorbed fraction.
  • 【Application】
    Purification of phosphorylated proteins in A431 lysate
    M:Molecular Marker
  • Lane 1:Non absorbed fraction
  • Lane 2:Absorbed fraction
  • Lane 3:Column rinsing fraction
  • (Left) Fluorescence staining
  • (right) Western blotting with anti-phosphorylated Tyr.