Rabbit MDM2 Polyclonal Antibody | anti-MDM2 antibody

Phospho-MDM2 (Tyr394) Antibody

Cat. Num. AAA9614385

• Gene Names: MDM2; HDMX; hdm2; ACTFS
• Reactivity: Human, Mouse, Monkey; Predicted Reactivity: Pig (100%), Zebrafish (100%), Bovine (100%), Horse (100%), Sheep (100%), Dog (100%), Chicken (100%), Xenopus (100%)
• Applications: Western Blot, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: MDM2; Polyclonal Antibody; Phospho-MDM2 (Tyr394) Antibody; ACTFS; Double minute 2 protein; E3 ubiquitin-protein ligase Mdm2; Hdm 2; Hdm2; HDMX; MDM 2; 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; anti-MDM2 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Monkey; Predicted Reactivity: Pig (100%), Zebrafish (100%), Bovine (100%), Horse (100%), Sheep (100%), Dog (100%), Chicken (100%), Xenopus (100%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-MDM2 (Tyr394) Antibody detects endogenous levels of MDM2 only when phosphorylated at Tyr394.; Tissue Specificity: 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.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-MDM2 antibody: Western Blot (WB), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IF/ICC: 1:100-1:500; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human MDM2 around the phosphorylation site of Tyr394.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: 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.
• Subunit Structure: Interacts with p53/TP53, TP73/p73, RBL5 and RP11. Binds specifically to RNA. Can interact with RB1, E1A-associated protein EP300 and the E2F1 transcription factor. Forms a ternary complex with p53/TP53 and WWOX. Interacts with CDKN2AIP, RFWD3, USP7, PYHIN1, and RBBP6. Interacts with ARRB1 and ARRB2. Interacts with PSMA3. Found in a trimeric complex with MDM2, MDM4 and USP2. Interacts with USP2 (via N-terminus and C-terminus). Interacts with MDM4. Part of a complex with MDM2, DAXX, RASSF1 and USP7. Part of a complex with DAXX, MDM2 and USP7. Interacts directly with DAXX and USP7. Interacts (via C-terminus) with RASSF1 isoform A (via N-terminus); the interaction is independent of TP53. Interacts with APEX1; leading to its ubiquitination and degradation. Interacts with RYBP; this inhibits ubiquitination of TP53. Identified in a complex with RYBP and p53/TP53. Also component of the TRIM28/KAP1-MDM2-p53/TP53 complex involved in regulating p53/TP53 stabilization and activity. Binds directly both p53/TP53 and TRIM28. Component of the TRIM28/KAP1-ERBB4-MDM2 complex involved in connecting growth factor responses with DNA damage. Interacts directly with both TRIM28 and ERBB4 in the complex. Interacts with DYRK2. Interacts with IGF1R. Interacts with TRIM13; the interaction ubiquitinates MDM2 leading to its proteasomal degradation. Interacts with SNAI1; this interaction promotes SNAI1 ubiquitination. Interacts with NOTCH1 (via intracellular domain). Interacts with FHIT. Interacts with RFFL and RNF34; the interaction stabilizes MDM2. Interacts with CDK5RAP3 and CDKN2A/ARF; form a ternary complex involved in regulation of p53/TP53. Interacts with MTA1. Interacts with AARB2. Interacts with MTBP. Interacts with PML. Interacts with TBRG1. Interacts with the 5S RNP which is composed of the 5S RNA, RPL5 and RPL11; the interaction is direct, occurs in the nucleoplasm and negatively regulates MDM2-mediated TP53 ubiquitination and degradation. Interacts with ADGRB1; the interaction results in inhibition of MDM2-mediated ubiquitination and degradation of DLG4/PSD95, promoting DLG4 stability and regulating synaptic plasticity (By similarity). Interacts with RPL23A; this interaction may promote p53/TP53 polyubiquitination. (Microbial infection) Interacts with herpes virus 8 protein v-IRF4. (Microbial infection) Interacts with and ubiquitinates HIV-1 Tat.
• Similarity: 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.
• Subcellular Location: 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.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of MDM2 (Phospho-Tyr394) using Insulin treated 293 whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunofluorescence (IF)/Immunocytochemistry (ICC)

(Staining COS7 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100,then blocked in 10% serum for 45 minutes at 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

Related Product Information for anti-MDM2 antibody

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).

NCBI and Uniprot Product Information

• NCBI GI #: 89993689
• NCBI GeneID: 4193
• NCBI Accession #: NP_002383.2
• NCBI GenBank Nucleotide #: NM_002392.5
• UniProt Accession #: Q00987; Q13226; Q13297; Q13298; Q13299; Q13300; Q13301; Q53XW0; Q71TW9; Q8WYJ1; A6NL51; A8K2S6
• Molecular Weight: 55,233 Da
• NCBI Official Full Name: E3 ubiquitin-protein ligase Mdm2 isoform a
• NCBI Official Synonym Full Names: MDM2 oncogene, E3 ubiquitin protein ligase
• NCBI Official Symbol: MDM2
• NCBI Official Synonym Symbols: HDMX; hdm2; ACTFS
• NCBI Protein Information: E3 ubiquitin-protein ligase Mdm2; oncoprotein Mdm2; Mdm2, p53 E3 ubiquitin protein ligase homolog; double minute 2, human homolog of; p53-binding protein; Mdm2, transformed 3T3 cell double minute 2, p53 binding protein
• UniProt Protein Name: E3 ubiquitin-protein ligase Mdm2
• Protein Family: Mdm2-binding protein
• UniProt Gene Name: MDM2
• UniProt Synonym Gene Names: Hdm2
• UniProt Entry Name: MDM2_HUMAN

NCBI Description

This gene encodes a nuclear-localized E3 ubiquitin ligase. The encoded protein can promote tumor formation by targeting tumor suppressor proteins, such as p53, for proteasomal degradation. This gene is itself transcriptionally-regulated by p53. Overexpression or amplification of this locus is detected in a variety of different cancers. There is a pseudogene for this gene on chromosome 2. Alternative splicing results in a multitude of transcript variants, many of which may be expressed only in tumor cells. [provided by RefSeq, Jun 2013]

Uniprot Description

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.

Protein type: Ligase; EC 6.3.2.19; EC 6.3.2.-; Ubiquitin ligase; Oncoprotein; Inhibitor; Ubiquitin conjugating system; Nucleolus

Chromosomal Location of Human Ortholog: 12q14.3-q15

Cellular Component: nucleoplasm; nuclear body; protein complex; cytoplasm; nucleolus; plasma membrane; synapse; cytosol; nucleus

Molecular Function: identical protein binding; protein binding; peroxisome proliferator activated receptor binding; enzyme binding; zinc ion binding; p53 binding; ubiquitin protein ligase binding; ubiquitin-protein ligase activity; ligase activity

Biological Process: viral reproduction; nerve growth factor receptor signaling pathway; protein ubiquitination during ubiquitin-dependent protein catabolic process; protein ubiquitination; response to morphine; negative regulation of caspase activity; negative regulation of transcription from RNA polymerase II promoter; positive regulation of mitotic cell cycle; DNA damage response, signal transduction by p53 class mediator resulting in cell cycle arrest; response to carbohydrate stimulus; response to antibiotic; positive regulation of protein export from nucleus; synaptic transmission; negative regulation of DNA damage response, signal transduction by p53 class mediator; response to magnesium ion; establishment of protein localization; peptidyl-lysine modification; positive regulation of cell proliferation; protein complex assembly; response to iron ion; epidermal growth factor receptor signaling pathway; response to drug; traversing start control point of mitotic cell cycle; phosphoinositide-mediated signaling; fibroblast growth factor receptor signaling pathway; protein destabilization; response to ether; response to cocaine; positive regulation of proteasomal ubiquitin-dependent protein catabolic process; innate immune response; regulation of protein catabolic process; negative regulation of transcription, DNA-dependent

Disease: Accelerated Tumor Formation, Susceptibility To