MDM2 Antibody - #AF0208

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製品: MDM2 Antibody
カタログ: AF0208
タンパク質の説明: Rabbit polyclonal antibody to MDM2
アプリケーション: WB IHC IF/ICC IP
Cited expt.: WB, IF/ICC
反応性: Human, Mouse, Rat, Monkey
予測: Pig, Bovine, Horse, Chicken
分子量: 45~55kD,70~95kD; 55kD(Calculated).
ユニプロット: Q00987
RRID: AB_2833395

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MS Report
HPLC Report
MSDS
説明書
COA
プロトコル
WB Handbook
IHC Protocol
IF/ICC Protocol

製品説明

ソース:
Rabbit
アプリケーション:
WB 1:500-1:3000, IHC 1:50-1:200, IF/ICC: 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

反応性:
Human,Mouse,Rat,Monkey
予測:
Pig(88%), Bovine(88%), Horse(88%), Chicken(86%)
クローナリティ:
Polyclonal
特異性:
MDM2 Antibody detects endogenous levels of total MDM2.
RRID:
AB_2833395
引用形式: Affinity Biosciences Cat# AF0208, RRID:AB_2833395.
コンジュゲート:
Unconjugated.
精製:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
保存:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
別名:

折りたたみ／展開

ACTFS; Double minute 2 protein; E3 ubiquitin-protein ligase Mdm2; Hdm 2; Hdm2; HDMX; MDM 2; MDM2; MDM2 oncogene E3 ubiquitin protein ligase; Mdm2 p53 E3 ubiquitin protein ligase homolog; Mdm2 transformed 3T3 cell double minute 2 p53 binding protein (mouse) binding protein 104kDa; MDM2_HUMAN; MDM2BP; Mouse Double Minute 2; MTBP; Murine Double Minute Chromosome 2; Oncoprotein Mdm2; p53 Binding Protein Mdm2; p53-binding protein Mdm2; Ubiquitin protein ligase E3 Mdm2; Ubiquitin protein ligase E3 Mdm2;

免疫原

免疫原:

A synthesized peptide derived from human MDM2, corresponding to a region within the internal amino acids.

Uniprot:

Q00987(MDM2_HUMAN) >>Visit HPA database.

遺伝子(ID):
MDM2(4193)
発現特異性:
Q00987 MDM2_HUMAN:

Ubiquitous. Isoform Mdm2-A, isoform Mdm2-B, isoform Mdm2-C, isoform Mdm2-D, isoform Mdm2-E, isoform Mdm2-F and isoform Mdm2-G are observed in a range of cancers but absent in normal tissues.

タンパク質の説明:
MDM2 a ubiquitin ligase for p53, plays a central role in regulation of the stability of p53 via Akt-mediated MDM2 phosphorylation. Phosphorylation of MDM2 increases its interaction with p300, providing a platform to allow the assembly of the protein complex necessary for MDM2-mediated ubiquitination and degradation of p53. Phosphorylation of MDM2 also blocks its binding to p19ARF, increasing the degradation of p53. Facilitates the nuclear export of p53 and targets it for proteasome-mediated proteolysis. Eight alternatively spliced isoforms have been reported.
タンパク質配列:
MCNTNMSVPTDGAVTTSQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSENRCHLEGGSDQKDLVQELQEEKPSSSHLVSRPSTSSRRRAISETEENSDELSGERQRKRHKSDSISLSFDESLALCVIREICCERSSSSESTGTPSNPDLDAGVSEHSGDWLDQDSVSDQFSVEFEVESLDSEDYSLSEEGQELSDEDDEVYQVTVYQAGESDTDSFEEDPEISLADYWKCTSCNEMNPPLPSHCNRCWALRENWLPEDKGKDKGEISEKAKLENSTQAEEGFDVPDCKKTIVNDSRESCVEENDDKITQASQSQESEDYSQPSTSSSIIYSSQEDVKEFEREETQDKEESVESSLPLNAIEPCVICQGRPKNGCIVHGKTGHLMACFTCAKKLKKRNKPCPVCRQPIQMIVLTYFP

種類予測

種類予測:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
88
Horse
88
Bovine
88
Chicken
86
Dog
75
Sheep
0
Xenopus
0
Zebrafish
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

研究背景

機能:

E3 ubiquitin-protein ligase that mediates ubiquitination of p53/TP53, leading to its degradation by the proteasome. Inhibits p53/TP53- and p73/TP73-mediated cell cycle arrest and apoptosis by binding its transcriptional activation domain. Also acts as a ubiquitin ligase E3 toward itself and ARRB1. Permits the nuclear export of p53/TP53. Promotes proteasome-dependent ubiquitin-independent degradation of retinoblastoma RB1 protein. Inhibits DAXX-mediated apoptosis by inducing its ubiquitination and degradation. Component of the TRIM28/KAP1-MDM2-p53/TP53 complex involved in stabilizing p53/TP53. Also component of the TRIM28/KAP1-ERBB4-MDM2 complex which links growth factor and DNA damage response pathways. Mediates ubiquitination and subsequent proteasome degradation of DYRK2 in nucleus. Ubiquitinates IGF1R and SNAI1 and promotes them to proteasomal degradation. Ubiquitinates DCX, leading to DCX degradation and reduction of the dendritic spine density of olfactory bulb granule cells (By similarity). Ubiquitinates DLG4, leading to proteasomal degradation of DLG4 which is required for AMPA receptor endocytosis (By similarity).

PTMs:

Phosphorylation on Ser-166 by SGK1 activates ubiquitination of p53/TP53. Phosphorylated at multiple sites near the RING domain by ATM upon DNA damage; this prevents oligomerization and E3 ligase processivity and impedes constitutive p53/TP53 degradation.

Autoubiquitination leads to proteasomal degradation; resulting in p53/TP53 activation it may be regulated by SFN. Also ubiquitinated by TRIM13. Deubiquitinated by USP2 leads to its accumulation and increases deubiquitination and degradation of p53/TP53. Deubiquitinated by USP7 leading to its stabilization.

細胞の位置付け:

Nucleus>Nucleoplasm. Cytoplasm. Nucleus>Nucleolus.
Note: Expressed predominantly in the nucleoplasm. Interaction with ARF(P14) results in the localization of both proteins to the nucleolus. The nucleolar localization signals in both ARF(P14) and MDM2 may be necessary to allow efficient nucleolar localization of both proteins. Colocalizes with RASSF1 isoform A in the nucleus.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
組織特異性:

Ubiquitous. Isoform Mdm2-A, isoform Mdm2-B, isoform Mdm2-C, isoform Mdm2-D, isoform Mdm2-E, isoform Mdm2-F and isoform Mdm2-G are observed in a range of cancers but absent in normal tissues.

タンパク質ファミリー:

Region I is sufficient for binding p53 and inhibiting its G1 arrest and apoptosis functions. It also binds p73 and E2F1. Region II contains most of a central acidic region required for interaction with ribosomal protein L5 and a putative C4-type zinc finger. The RING finger domain which coordinates two molecules of zinc interacts specifically with RNA whether or not zinc is present and mediates the heterooligomerization with MDM4. It is also essential for its ubiquitin ligase E3 activity toward p53 and itself.

Belongs to the MDM2/MDM4 family.

研究領域

· Cellular Processes > Cell growth and death > Cell cycle.   (View pathway)

· Cellular Processes > Cell growth and death > p53 signaling pathway.   (View pathway)

· Cellular Processes > Transport and catabolism > Endocytosis.   (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence.   (View pathway)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Genetic Information Processing > Folding, sorting and degradation > Ubiquitin mediated proteolysis.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Drug resistance: Antineoplastic > Platinum drug resistance.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Infectious diseases: Viral > Epstein-Barr virus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Transcriptional misregulation in cancer.

· Human Diseases > Cancers: Overview > Viral carcinogenesis.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Overview > MicroRNAs in cancer.

· Human Diseases > Cancers: Specific types > Glioma.   (View pathway)

· Human Diseases > Cancers: Specific types > Prostate cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Melanoma.   (View pathway)

· Human Diseases > Cancers: Specific types > Bladder cancer.   (View pathway)

· Human Diseases > Cancers: Specific types > Chronic myeloid leukemia.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

参考文献

1). Regorafenib inhibits EphA2 phosphorylation and leads to liver damage via the ERK/MDM2/p53 axis. Nature Communications, 2023 (PubMed: 37179400) [IF=16.6]

Application: WB    Species: Human    Sample: HL-7702 cells

Fig. 5 The phosphorylation of EphA2Ser897 regulates p53 by ERK/MDM2 axis. a HL-7702 cells were treated with regorafenib as indicated. The expression levels of p-MDM2 (Ser166) and MDM2 were analyzed by western blot. n = 4 independent experiments. b C57BL/6J mice were treated with or without 400 mg/kg/day regorafenib for 6 weeks. The expression levels of p-MDM2 (Ser166) and p53 in liver tissues were analyzed by western blot (n = 5 per group). c HL-7702 cells treated with 8 μM regorafenib for 24 h (n = 3 independent experiments) or liver tissues (n = 3 per group) were used to detect the expression level of Lamin B1 and MDM2 in whole cell, cytoplasm and nuclei by western blot. d, e HL-7702 cells transfected with vector, EphA2 S897A plasmid or EphA2 S897D plasmid as indicated for 24 h. d The expression levels of HA, p-MDM2 (Ser166) and p53 were analyzed by western blot. e The localization of MDM2 was observed by immunofluorescence. Representative images are shown from three independent experiments. Scale bar: 20 µm. f HL-7702 cells were treated with ALW-II-41-27 or bosutinib as indicated for 24 h. Related proteins were analyzed by western blot. Blots are representative of three independent experiments. g HL-7702 cells were treated with regorafenib as indicated for 24 h. The expression levels of related proteins were analyzed by western blot. The relative level of p-ERK (Tyr204) was analyzed by densitometric analysis. n = 3 independent experiments. h HL-7702 cells were treated with regorafenib as indicated. The expression levels of related proteins were analyzed by western blot. The fold change of pS-EphA2/EphA2 and pY-ERK/ERK were analyzed by densitometric analysis. n = 3 independent experiments. Data were expressed as mean ± SD. Unpaired two-sided Student’s t test for b and c or one way ANOVA followed by Tukey post hoc test for a and g. Source data are provided as a Source Data file. Regora regorafenib, MW molecular weight, WCL whole cell lysate, CE cytoplasmic extraction, NE nuclear extraction.

Application: IF/ICC    Species: Human    Sample: HL-7702 cells

Fig. 5 The phosphorylation of EphA2Ser897 regulates p53 by ERK/MDM2 axis. a HL-7702 cells were treated with regorafenib as indicated. The expression levels of p-MDM2 (Ser166) and MDM2 were analyzed by western blot. n = 4 independent experiments. b C57BL/6J mice were treated with or without 400 mg/kg/day regorafenib for 6 weeks. The expression levels of p-MDM2 (Ser166) and p53 in liver tissues were analyzed by western blot (n = 5 per group). c HL-7702 cells treated with 8 μM regorafenib for 24 h (n = 3 independent experiments) or liver tissues (n = 3 per group) were used to detect the expression level of Lamin B1 and MDM2 in whole cell, cytoplasm and nuclei by western blot. d, e HL-7702 cells transfected with vector, EphA2 S897A plasmid or EphA2 S897D plasmid as indicated for 24 h. d The expression levels of HA, p-MDM2 (Ser166) and p53 were analyzed by western blot. e The localization of MDM2 was observed by immunofluorescence. Representative images are shown from three independent experiments. Scale bar: 20 µm. f HL-7702 cells were treated with ALW-II-41-27 or bosutinib as indicated for 24 h. Related proteins were analyzed by western blot. Blots are representative of three independent experiments. g HL-7702 cells were treated with regorafenib as indicated for 24 h. The expression levels of related proteins were analyzed by western blot. The relative level of p-ERK (Tyr204) was analyzed by densitometric analysis. n = 3 independent experiments. h HL-7702 cells were treated with regorafenib as indicated. The expression levels of related proteins were analyzed by western blot. The fold change of pS-EphA2/EphA2 and pY-ERK/ERK were analyzed by densitometric analysis. n = 3 independent experiments. Data were expressed as mean ± SD. Unpaired two-sided Student’s t test for b and c or one way ANOVA followed by Tukey post hoc test for a and g. Source data are provided as a Source Data file. Regora regorafenib, MW molecular weight, WCL whole cell lysate, CE cytoplasmic extraction, NE nuclear extraction.
2). Pimozide inhibits the growth of breast cancer cells by alleviating the Warburg effect through the P53 signaling pathway. BIOMEDICINE & PHARMACOTHERAPY, 2022 (PubMed: 35658233) [IF=6.9]

Application: WB    Species: Human    Sample: MCF-7 cells

Fig. 5. Pimozide promotes the expression of p53 through PI3K/Akt/MDM2 pathway. (A-B) Cells were treated with the indicated concentrations of Pimozide for 24 h, the protein expression of p-PI3K, PI3K, p-Akt, Akt, p-MDM2, MDM2 in MCF-7 (A) and MDA-MB-231 (B) cells were determined by Western blot analysis (left panel). Densitometry analysis was performed to assess the protein expression of p-PI3K/PI3K, p-Akt/Akt, p-MDM2/MDM2 (normalized to β-actin expression) (right panel). (C-F) Western blot analysis for p-PI3K, PI3K, p-Akt, Akt, p-MDM2, MDM2, P53, and PKM2 protein levels in MCF-7 (C, E) and MDA-MB-231 (D, F) cells incubated with PI3K inhibitor LY294002 or PI3K agonist SC79 as indicated. (G) After treatment with SC79, colony-forming assay images (left panel) and quantification of colony number percentages (right panel). Data represent mean ± SD from three biological replicates (*p < 0.05, **p < 0.01, ns: not significant).
3). Notoginsenoside R1 attenuates ischemic heart failure by modulating MDM2/β arrestin2-mediated β2-adrenergic receptor ubiquitination. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie, 2024 (PubMed: 38955084) [IF=6.9]

Application: WB    Species: Mouse    Sample:

Fig. 6. β2AR ubiquitanation was inhibited by NGR1 via regulation of the MDM2/β-arrestin2 complex. (A) The impact of NGR1 on the levels of β-arrestin2 and MDM2 in the cardiac of mice with CHF. Results were depicted as Mean ± SEM (n=5). To compare the various groups statistically, a one-way nested ANOVA followed by Dunnett's test was conducted. #P < 0.05, ##P < 0.01, the Sham vs. the CHF group; *P < 0.05, **P < 0.01, ***P < 0.001, the NGR1 groups vs. the CHF group. (B) MDM2 and β-arrestin2 expression in H9c2 cells stimulated with OGD in response to NGR1. Results are depicted as Mean ± SEM (n = 3). To compare the various groups statistically, a one-way nested ANOVA followed by Dunnett's test #P < 0.05, ##P < 0.01, the control vs. the OGD group; *P < 0.05, **P < 0.01, ***P < 0.001, the NGR1 groups vs. the OGD group. (C) The influence of NGR1 on the β-arrestin2–MDM2 interaction. (D) The impact of NGR1 in the presence of SP141 on the viability of H9c2 cells after OGD/R damage. Following a one-hour pretreatment with SP141, the cells were exposed to OGD for six hours before being treated with NGR1 for an additional twenty-four hours. Results are depicted as Mean ± SEM (n=3), ###P < 0.001, the control vs. the OGD group; ***P < 0.001, the NGR1 group vs. the OGD group; n.s. indicates no statistical significance.
4). Xinbao Pill attenuated chronic heart failure by suppressing the ubiquitination of β-adrenergic receptors. Phytomedicine, 2023 (PubMed: 37149964) [IF=6.7]
5). Terazosin, a repurposed GPR119 agonist, ameliorates mitophagy and β-cell function in NAFPD by inhibiting MST1-Foxo3a signalling pathway. Cell proliferation, 2024 (PubMed: 39413003) [IF=5.9]
6). Storax Attenuates Cardiac Fibrosis following Acute Myocardial Infarction in Rats via Suppression of AT1R–Ankrd1–P53 Signaling Pathway. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2022 (PubMed: 36361958) [IF=5.6]

Application: WB    Species: Rat    Sample: cardiac tissue

Figure 8 Storax inhibits the AT1R–Ankrd1–P53 pathway in AMI rats against cardiomyocyte apoptosis. (A) WB bands showing the protein expression levels of AT1R, Ankrd1, P-p53 (ser15), P53 and Mdm2 in cardiac tissue. (B–E) Relative protein for AT1R, Ankrd1, P-p53 (ser15) and Mdm2 were quantified by densitometry based on immunoblot images. Results are presented as mean ± SD (n = 3). One-way ANOVA follow by Bonferroni’s post hoc test: # p < 0.05, ## p < 0.01 vs. vehicle group.
7). Dehydrocorydaline attenuates myocardial ischaemia-reperfusion injury via the FoXO signalling pathway: a multimodal study based on network pharmacology, molecular docking, and experimental study. Journal of ethnopharmacology, 2024 (PubMed: 39222757) [IF=4.8]

Application: WB    Species: Rat    Sample: H9c2 cells

Fig. 7. Effects of DHC on the expression of predicted targets in the in vitro model of H/R injury. (A) Representative fluorescence images of the TUNEL assay. Photographs were taken at × 400 magnification. (B) Percentage of TUNEL-positive cells in each group. (C) Western blot showing the protein expression of cleaved-caspase 3 and cleaved-caspase 8 in each group. (D) Western blot showing the protein expression of p-FOXO1A, FOXO1A, CCND1, p-MDM2, and MDM2 in each group. (E) Relative protein levels of cleaved-caspase 3 and cleaved-caspase 8 measured in western blots (n = 3). (F) Relative protein levels of p-FOXO1A, FOXO1A, and p-FOXO1A/FOXO1A measured in western blots (n = 3). (G) Relative protein levels of p-MDM2, MDM2, and p-MDM2/MDM2 measured in western blots (n = 3). (H) Relative protein levels of CCND1 measured in western blots (n = 3). * indicates a significant difference compared to the control group; # indicates a significant difference compared to the H/R model group: *p < 0.05, **p < 0.01, #p < 0.05, ##p < 0.01. DHC: dehydrocorydaline, H/R: hypoxia/reoxygenation, TUNEL: TdT-mediated dUTP-biotin nick end labelling.
8). Dehydrocorydaline attenuates myocardial ischemia-reperfusion injury via the FoXO signalling pathway: A multimodal study based on network pharmacology, molecular docking, and experimental study. Journal of ethnopharmacology, 2024 (PubMed: 39222757) [IF=4.8]
9). Casein kinase 1α inhibits p53 downstream of MDM2‑mediated autophagy and apoptosis in acute myeloid leukemia. ONCOLOGY REPORTS, 2020 (PubMed: 32901886) [IF=3.8]

Application: WB    Species: human    Sample: HEL cells.

Figure 5. |Autophagy inhibitor Spautin-1 aggravates apoptosis induced by D4476, and CK1α inhibits autophagy by targeting the p53/AMPK/mTOR signaling pathway downstream of MDM2 in AML cells. (C) Co‑immunoprecipitation showed that CK1α interacted with MDM2 and p53 in HEL cells. Ten percent whole cell lysate was used as input samples.
10). Linc-ROR promotes arsenite-transformed keratinocyte proliferation by inhibiting P53 activity. Metallomics, 2020 (PubMed: 32373892) [IF=2.9]

Application: WB    Species: human    Sample: HaCaT cells

Fig. 3 | P53 activity was decreased with arsenite-induced transformation of HaCaT cells. HaCaT cells were continuously maintained in medium containing 1 mM sodium arsenite for 0, 20, 35 and 50 passages. (a) The relative level of P53 mRNA in 0, 20, 35 and 50 passages of cells, with the control group set to 1 (the same hereinafter). (b–d) The western blotting of P53 and MDM2 proteins; GAPDH served as the loading control.

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