MDM2/HDM2 Antibody (A10E11-R)

Western Blot: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Western blot analysis of MDM2/HDM2 on different lysates with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) at 1/1,000 dilution. Lane 1: U-2 OS cell lysate Lane 2: U-2 OS treated with 10uM NUTLIN 3A for 24 hours cell lysate Lysates/proteins at 15 ug/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 (NBP3-32584) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Immunohistochemical analysis of paraffin-embedded human embryonal carcinoma tissue with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) 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 (NBP3-32584) 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.

Immunocytochemistry/ Immunofluorescence: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Immunocytochemistry analysis of U-2 OS cells treated with or without 10μM NUTLIN 3A for 24 hours labeling MDM2/HDM2 with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) 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/HDM2 antibody (NBP3-32584) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.

• Reactivity: Hu, Pm
• Applications: WB, ICC/IF, IHC
• Clone: A10E11-R
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

MDM2/HDM2 Antibody (A10E11-R) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Recombinant protein within human MDM2/HDM2 aa 1-491 / 491. (Uniprot: Q00987)
• Localization: Nucleus, nucleoplasm, nucleolus, Cytoplasm.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: MDM2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin 1:1000
  • Western Blot 1:1000
• Theoretical MW: 55 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS (pH7.4), 0.1% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for MDM2/HDM2 Antibody (A10E11-R)

• ACTFS
• HDM2
• HDMX
• MDM2 oncogene, E3 ubiquitin protein ligase
• MDM2

Background

MDM2 is a target gene of the transcription factor tumor protein p53. The encoded protein is a nuclear phosphoprotein that binds and inhibits transactivation by tumor protein p53, as part of an autoregulatory negative feedback loop. Overexpression of this gene can result in excessive inactivation of tumor protein p53, diminishing its tumor suppressor function. This protein has E3 ubiquitin ligase activity, which targets tumor protein p53 for proteasomal degradation. This protein also affects the cell cycle, apoptosis, and tumorigenesis through interactions with other proteins, including retinoblastoma 1 and ribosomal protein L5. More than 40 different alternatively spliced transcript variants have been isolated from both tumor and normal tissues.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: MDM2