Alexa Fluor® 647 Anti-MDM2 antibody [SMP 14] (ab216076)

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-MDM2 antibody [SMP 14] (ab216076)

    Key features and details

    • Alexa Fluor® 647 Mouse monoclonal [SMP 14] to MDM2
    • Suitable for: ICC/IF
    • Reacts with: Human
    • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
    • Isotype: IgG1

    Overview

    • Product name

      Alexa Fluor® 647 Anti-MDM2 antibody [SMP 14]
      See all MDM2 primary antibodies

    • Description

      Alexa Fluor® 647 Mouse monoclonal [SMP 14] to MDM2

    • Host species

      Mouse

    • Conjugation

      Alexa Fluor® 647. Ex: 652nm, Em: 668nm

    • Specificity

      This antibody also recognises a peptide epitope around Thr-216 of murine MDM2 (when Thr-216 is unphosphorylated). SMP14 also cross reacts with some cytokeratins (6, 14 & 16). This is only a problem when working with certain epithelial cells and not fibroblasts.

    • Tested applications

      Suitable for: ICC/IFmore details

    • Species reactivity

      Reacts with: Human
      Predicted to work with: Mouse, Rat

    • Immunogen

      Synthetic peptide corresponding to Human MDM2 aa 150-250 conjugated to keyhole limpet haemocyanin (Glutaraldehyde).
      Database link: Q00987

    • Epitope

      Amino acids 154 - 167.

    • Positive control

      • ICC/IF: HeLa cells

    • General notes

      Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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    Properties

    Applications

    Our Abpromise guarantee covers the use of ab216076 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    ICC/IF 1/1000.

    This product gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)

    Target

    • Function

      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 an 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.

    • 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.

    • Involvement in disease

      Note=Seems to be amplified in certain tumors (including soft tissue sarcomas, osteosarcomas and gliomas). A higher frequency of splice variants lacking p53 binding domain sequences was found in late-stage and high-grade ovarian and bladder carcinomas. Four of the splice variants show loss of p53 binding.

    • Sequence similarities

      Belongs to the MDM2/MDM4 family.
      Contains 1 RanBP2-type zinc finger.
      Contains 1 RING-type zinc finger.
      Contains 1 SWIB domain.

    • Domain

      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.

    • Post-translational
      modifications

      Phosphorylated in response to ionizing radiation in an ATM-dependent manner.
      Auto-ubiquitinated; which leads to proteasomal degradation. Deubiquitinated by USP2 leads to its accumulation and increases deubiquitinilation and degradation of p53/TP53. Deubiquitinated by USP7; leading to stabilize it.

    • Cellular localization

      Nucleus > nucleoplasm. Cytoplasm. Nucleus > nucleolus. 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.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

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

    Images

    • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-MDM2 antibody [SMP 14] (ab216076)
      Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-MDM2 antibody [SMP 14] (ab216076)

      Ab216076 staining MDM2 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab216076 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

       

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    References (0)

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    ab216076 has not yet been referenced specifically in any publications.

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