Anti-cMyc phospho-Ser62 antibody, mouse monoclonal (33A12E10), Validated
Product code

71-161Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol

PurityPurified IgG fraction with protein A from hybridoma cell culture medium.

ImmunogenSynthetic peptide containing phospho-Ser62 of cMyc

Isotype

Mouse IgG2b κReactivity

Human and mouse.

Special notes

Specificity of this antibody have been independently validated by siRNAApplication

1. Western blotting (~1µg/ml, Fig.1)
2. Immunofluorescence staining (0.25~1 μg/ml)
3. Immunohistochemistry (5 μg/ml), Paraffin-embedded
4. Flow cytometry (1 μg for 106 cells)
5. Indirect ELISA

Background(Assay dependent concentration)

cMyc is a proto-oncogene, which is overexpressed in a wide range of humancancers.

Myc gene encodes a transcription factor that regulates a great

number of genes through binding on Enhancer Box sequences (E-boxes) andrecruiting histone acetyltransferase.

It can also act as a transcriptional

repressor. It regulates cell growth, apoptosis, differentiation and stem cell selfrenewal.

Previous studies on the phosphorylation of c-Myc have suggested

functional association between phosphorylation at Thr58/Ser62 by glycogensynthase kinase 3, cyclin dependent kinase, ERK2 and C-Jun N terminal Kinase(JNK), cell proliferation and cell cycle regulation. Phosphorylation at Ser62 isrequired for Ras-induced stabilization and is prerequisite for phosphorylation
at Thr58 for its degradation (ref.1).

Data Link

UniProtKB/Swiss-Prot P01106 (MYC_HUMAN)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
1 / 3

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Data Images: 71-161 Anti-cMyc phospho-Ser62 antibody, mouse monoclonal (33A12E10), Validated←cMyc p-S62

Fig.1. Identification of cMyc protein whose Ser62 is phosphorylated by Western blotting.
Samples:Crude cell extracts of AGS (gastric adenocarcinoma) cells.
Scr; Scrambled siRNA was introduced into the cells as a negative control.
Neg.Contol; Negative control siRNA was transfected.
Myc1; siRNA for cMyc was transfected. (The data was provided by Drs.A. Khanna and J.Westermarckof University of Tampere)

Fig.2. Immunofluorescence staining of cMyc phosho-Ser62 in HeLa cells
Left: Cells stained with anti-cMyc pS62 antibody (green) and DAPI (blue)
Right: Proximity Ligation Analysis with anti-cMyc pS62 and CIP2A antibodies,
association of cMyc pS62 with CIP2A (red) in nuclei (DAPI, blue)
Images kindly provided by Prof Westermarck J and Dr. Qiao X. For details refer to Ref.1
2 / 3

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Anti-cMyc p62 antibody

DAPIMerge

Fig.3. Immunofluorescence staining of cMyc phosho-Ser62 in nuclei of HeLa cells.
1. HeLa cells were fixed with 4% paraformaldehyde overnight, permealized with 0.25% Triton
X-100 in PBS for 10 min.
2. Incubate cells with 1.5% BSA in PBS for 30 min to block non-specific binding of theantibodies. Incubate the cells with 1/4,000 diluted anti-cMyc p62 antibody in 1% BSA inPBS at 4℃ overnight.
3. Incubate cells with a secondary antibody, goat anti-mouse IgG conjugated with Alex 488, at
1/1,000 dilution in 1% BSA for 1 hr at room temperature.
4. Nucleus (DNA) was stained with DAPI
References: This product was used in the following Publications.
1. Myant K et al. Serine 62-Phosphorylated MYC Associates with Nuclear Lamins and Its
Regulation by CIP2A Is Essential for Regenerative Proliferation. Cell Rep. 2015 Aug
11;12(6):1019-31. PMID: 26235622. WB, IF (human, mouse)
2.

Tibbitts DC et al. Studying c-Myc serine 62 phosphorylation in leukemia cells: concern overantibody cross-reactivity. Blood 119:5334-5 (2012).PubMed: 22653959 WB, IP (human)

3. Mathiasen DP. Identification of a c-Jun N-terminal kinase-2-dependent signal amplificationcascade that regulates c-Myc levels in ras transformation. Oncogene. 2012 Jan 19;31(3):390-401.
PubMed: 21706057 WB (mouse)
4. Wang X. et al. Phosphorylation regulates c-Myc’s oncogenic activity in the mammary gland. Cancer
Res. 2011 Feb 1; 71(3): 925–936. PubMed: 3077809 WB (human)
5. Khanna A. MYC-dependent regulation and prognostic role of CIP2A in gastric cancer. J Natl Cancer Inst. 2009
Jun 3;101(11):793-805. PubMed: 19470954 WB (human)

3 / 3

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/