Anti-MDM2 antibody
cat.: ER1902-14
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, ICC, IHC-P, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1 mg/mL.
Purification: Protein A affinity purified.
Molecular weight: 55 kDa
Isotype: IgG
Immunogen: Recombinant protein within human MDM2 aa 1-100.
Positive control: Human lung tissue lysate, Daudi cell lysate, HepG2, JAR, MCF-7, human thyroid tissue, human skin tissue, human breast carcinoma tissue, human small intestine tissue, human pancreas tissue, Daudi.
Subcellular location: Nucleoplasm, nucleolus, cytoplasm.
Recommended Dilutions:
  WB
  ICC
  IHC-P
  FC

1:500-1:1,000
1:50-1:200
1:500-1:1,000
1:50-1:100
Uniprot #: SwissProt: Q00987 Human
Alternative names: 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
Images
ER1902-14_1.jpg Fig1: Western blot analysis of MDM2 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: human lung tissue lysate
Lane 2: Daudi cell lysate
ER1902-14_2.jpg Fig2: ICC staining of MDM2 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1902-14, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1902-14_3.jpg Fig3: ICC staining of MDM2 in JAR cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1902-14, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1902-14_4.jpg Fig4: ICC staining of MDM2 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ER1902-14, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ER1902-14_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-MDM2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-14, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1902-14_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-MDM2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-14, 1/500) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1902-14_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-MDM2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-14, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1902-14_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-MDM2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-14, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1902-14_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-MDM2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER1902-14, 1/1,000) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ER1902-14_10.jpg Fig10: Flow cytometric analysis of MDM2 was done on Daudi cells. The cells were fixed, permeabilized and stained with the primary antibody (ER1902-14, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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