Phos-tag SDS-PAGE database　53BP1

53BP1

Double-strand break (DSB) repair protein involved in response to DNA damage, telomere dynamics and class-switch recombination (CSR) during antibody genesis
Mw; 213,000

Image

53BP1

53BP1 is a high molecular protein, so the protein was analyzed by 3%(w/v) polyacrylamidegel strengthened by 0.5%(w/v) agarose gel.

Upper panels are normal SDS-PAGE, and lower panels are Mn²⁺–Phos-tag SDS-PAGE.

HeLa cells were cultured in DMEM without serum, and treated actinomycinD (2µM) for 5 hrs.
Cells were lyzed by SDS-PAGE loading buffer.

Reference	Kinoshita, E., Kinoshita-Kikuta, E., Ujihara, H., and Koike, T. Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agarose. *Proteomics, 9, 4098–4101 (2009) doi: 10.1002/pmic.200900020. 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.
Composition of gel	Mn²⁺–Phos-tag:　20µM Polyacrylamide: 3%(w/v) + Agarose 0.5%(w/v)
Detection	WB: anti-53BP1 anti-p53BP1(pS25/29) anti-p53BP1(pS1778)

Reference

Kinoshita, E., Kinoshita-Kikuta, E., Ujihara, H., and Koike, T.
Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agarose.
Proteomics, 9, 4098–4101 (2009)

doi: 10.1002/pmic.200900020.

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.

Composition of gel

Mn²⁺–Phos-tag:　20µM

Polyacrylamide: 3%(w/v)
+
Agarose 0.5%(w/v)

Detection

WB:

anti-53BP1

anti-p53BP1(pS25/29)

anti-p53BP1(pS1778)