Phos-tag SDS-PAGE database EGFR

EGFR

Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses.
Mw; 134,000

Image

EGFR is a high molecular protein, so the protein was analyzed by polyacrylamide gel with low concentration.

HeLa cells were treated with EGF for 5 min and then cells were lyzed by SDS-PAGE loading buffer.

Phosphorylation of EGFR was analyzed by 5% or 3.5% (strengthened wit 0.5% agarose).

Related data

EGFR( Zn²⁺–Phos-tag SDS-PAGE)

Reference	Kinoshita, E., Kinoshita-Kikuta, E., and Koike, T. Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE. Nature Protocols, 9, 1513–1521 (2009) doi: 10.1038/nprot.2009.154 Supplemental data
Composition of gel	1. Mn²⁺–Phos-tag:　25µM Polyacrylamide: 5%(w/v) 2. Mn²⁺–Phos-tag:　25µM Polyacrylamide: 3.5%(w/v) + Agarose 0.5%(w/v)
Detection	WB: anti-EGFR

Reference

Kinoshita, E., Kinoshita-Kikuta, E., and Koike, T.
Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE.
Nature Protocols, 9, 1513–1521 (2009)

doi: 10.1038/nprot.2009.154

Supplemental data

Composition of gel

Mn²⁺–Phos-tag:　25µM

Polyacrylamide: 5%(w/v)

Mn²⁺–Phos-tag:　25µM

Polyacrylamide: 3.5%(w/v)
+
Agarose 0.5%(w/v)

Detection

　WB: anti-EGFR