MDM2 Recombinant Mouse Monoclonal Antibody [A10E11-R]
cat.: HA601310
| Product Type: | Recombinant Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Human, Monkey |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | A10E11-R |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 55 kDa |
| Isotype: | IgG1 |
| Immunogen: | Recombinant protein within human MDM2 aa 1-491 / 491. |
| Positive control: | U-2 OS treated with 10μM NUTLIN 3A for 24 hours cell lysate, HepG2 cell lysate, COS-1cell lysate, U-2 OS cells treated with 10μM NUTLIN 3A for 24 hours, human embryonal carcinoma tissue, human testis tissue. |
| Subcellular location: | Nucleus, nucleoplasm, nucleolus, Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:1,000 1:500 1:1,000 |
| 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
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Fig1:
Western blot analysis of MDM2 on different lysates with Mouse anti-MDM2 antibody (HA601310) at 1/1,000 dilution. Lane 1: U-2 OS cell lysate Lane 2: U-2 OS treated with 10μM NUTLIN 3A for 24 hours cell lysate Lysates/proteins at 15 µg/Lane. Predicted band size: 55 kDa Observed band size: 90 kDa Exposure time: 3 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601310) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of MDM2 on different lysates with Mouse anti-MDM2 antibody (HA601310) at 1/1,000 dilution. Lane 1: HepG2 cell lysate Lane 2: COS-1cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 55 kDa Observed band size: 90 kDa Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601310) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of U-2 OS cells treated with or without 10μM NUTLIN 3A for 24 hours labeling MDM2 with Mouse anti-MDM2 antibody (HA601310) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MDM2 antibody (HA601310) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human embryonal carcinoma tissue with Mouse anti-MDM2 antibody (HA601310) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601310) at 1/1,000 dilution for 1 hour 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. |
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Fig5:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Mouse anti-MDM2 antibody (HA601310) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601310) at 1/1,000 dilution for 1 hour 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. |
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