Anti-MDM2 (phospho S166) antibody [EPR1450(2)]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling MDM2 (phospho S166) using ab170880 at a 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

All lanes:

Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/50000 dilution

Lane 1:

MCF7 cell lysate at 10 µg

Lane 2:

MCF7 cell lysate treated with IGF-1 at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 55 kDa,62 kDa

Observed band size: 55 kDa

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• WB

Lab

Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

Diluting and blocking buffer : 2% BSA/TBST

The molecular weight is the same with the one from this paper PMID : 25392082

All lanes:

Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (ab170880) at 1/100000 dilution

Lane 1:

A549 (Human lung carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysate at 15 µg

Lane 3:

A549 (Human lung carcinoma epithelial cell) treated with insulin at 1ug/ml for 150 minutes. Whole cell lysate. Then the membrane was incubated with phosphatase at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 90-140 kDa

false

Exposure time: 5s

• Dot

Supplier Data

Dot Blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

Dot blot analysis of MDM2 (phospho S166) phospho peptide (Lane 1) MDM2 non-phospho peptide (Lane 2) labeling MDM2 (phospho S166) with ab170880 at a dilution of 1/1000. Goat Anti-Rabbit IgG (H+L) Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

• WB

CiteAb

Western blot - Anti-MDM2 (phospho S166) antibody [EPR1450(2)] (AB170880)

MDM2 (phospho S166) western blot using anti-MDM2 (phospho S166) antibody [EPR1450(2)] ab170880. Publication image and figure legend from Kou, F., Sun, H., et al., 2020, J Cancer, PubMed 32201520.

ab170880 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab170880 please see the product overview.

TOP2A is involved in p53 pathway. A. By analyzing GEO and TCGA databases, 148 mRNAs were found in TOP2A high and low groups. KEGG pathway enrichment analysis showed the top 3 most enriched pathways were Oocyte meiosis, p53 signaling pathway and Viral carcinogenesis, respectively. B. The heat map indicated significantly differentially expressed genes of P53 pathway between TOP2A-high and TOP2A-low group. CCNB1, CCNE2, CDK1, CCNB2, RRM2, CHEK1 and PMAIP1 were significantly upregulated in TOP2A high expression group. C. The expression levels of CCNB1, CCNB2 and CHEK1 were further identified by the violet plots, which demonstrated these genes expression levels were positively related to TOP2A. D. The expression levels of p53 related genes (cyclinB1 (CCNB1), cyclinB2 (CCNB2), CHEk1, MDM2, p53) in GLC82 and A549 cells with transfected TOP2A siRNA or control were identified by Western blot. Downregulation of the expression levels of CCNB1, CCNB2 and CHEk1 was observed in lung adenocarcinoma cells with siRNA compared with control. There was no difference in the expression levels of p53 and MDM2 between siRNA and control groups. These results indicated that TOP2A regulates the expression levels of p53 pathway related genes.

false