MDM2 Monoclonal antibody, PBS Only

MDM2 Monoclonal Antibody for WB, IF, IHC, Indirect ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

Human, mouse, rat, rabbit

Applications

WB, IF, IHC, Indirect ELISA

Conjugate

Unconjugated

CloneNo.

2A6C9

Cat no : 66511-1-PBS

Synonyms

Double minute 2 protein, hdm2, HDMX, MDM2, Oncoprotein Mdm2, p53 binding protein Mdm2

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Validation Data Gallery

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  WB analysis of MCF-7 using 66511-1-Ig (same clone as 66511-1-PBS)

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis of rat brain using 66511-1-Ig (same clone as 66511-1-PBS)

  rat brain tissue were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis of U-251 using 66511-1-Ig (same clone as 66511-1-PBS)

  U-251 cells were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis using 66511-1-Ig (same clone as 66511-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis of MCF-7 using 66511-1-Ig (same clone as 66511-1-PBS)

  MCF-7 cells were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:4000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis of mouse brain using 66511-1-Ig (same clone as 66511-1-PBS)

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  WB analysis of A549 using 66511-1-Ig (same clone as 66511-1-PBS)

  A549 cells were subjected to SDS PAGE followed by western blot with 66511-1-Ig (MDM2 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 66511-1-Ig (same clone as 66511-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66511-1-Ig (MDM2 antibody) at dilution of 1:300 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human breast cancer using 66511-1-Ig (same clone as 66511-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66511-1-Ig (MDM2 antibody) at dilution of 1:300 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human liver cancer using 66511-1-Ig (same clone as 66511-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66511-1-Ig (MDM2 antibody) at dilution of 1:300 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  IHC staining of human liver cancer using 66511-1-Ig (same clone as 66511-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 66511-1-Ig (MDM2 antibody) at dilution of 1:300 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

• 

  IF Staining of HeLa using 66511-1-Ig (same clone as 66511-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using MDM2 antibody (66511-1-Ig, Clone: 2A6C9 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66511-1-PBS in a different storage buffer formulation.

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Recommended dilution

Application	Dilution
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

66511-1-PBS targets MDM2 in WB, IF, IHC, Indirect ELISA applications and shows reactivity with Human, mouse, rat, rabbit samples.

• Tested Reactivity: Human, mouse, rat, rabbit
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: Mdm2 p53 binding protein homolog (mouse)
• Calculated Molecular Weight: 55 kDa
• Observed Molecular Weight: 40-55 kDa
• GenBank Accession Number: NM_002392
• Gene Symbol: MDM2
• Gene ID (NCBI): 4193
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS Only
• Storage Conditions: Store at -80°C.

Background Information

What is the cellular localization of MDM2?

MDM2 is predominantly localized in the nucleoplasm.

What is the tissue specificity of MDM2?

MDM2 is non-specific, expressing ubiquitously in the brain, heart, kidney, skin, and other tissues. Certain 

isoforms only express in cancerous cells and not normal tissue.

With what other molecules does MDM2 interact?

Most notably, the oncogene protein MDM2 regulates transcription factor p53 by binding its transcriptional 

activation domain and by E3-ubiquitin ligase activity. Ribosome proteins rpL5, rpL11, and rpL23 also bind to 

MDM2, inhibiting E3 ubiquitin ligase activity and promoting the p53 pathway.

What competes with MDM2 activity?

Nutilin-3 was designed to compete with MDM2 in binding with p53, inducing activation of the p53 pathway.

What is the role of ubiquitination in MDM2 activity?

While p53 is also affected by other post-translational modifications, its stability is determined by ubiquitination 

mediation of MDM2, suppressing p53 transcriptional regulation and leading to the proteosomal degradation of 

p53.

How is MDM2 associated with cancer?

MDM2 inhibits the activity of the tumor-suppressive protein p53, which serves as an important factor in 

promoting DNA repair or activating apoptotic pathways. Loss of p53 is associated with tumor development 

and growth.

What is the human ortholog of MDM2?

Human double-minute type 2 (HDM2)

In what cellular circumstances does the p53-MDM2 complex dissociate?

Cellular stress or DNA damage result in the phosphorylation and activation of p53. Phosphorylation also 

prevents p53-MDM2 interaction.

Where is the gene located?

Hdm2 is located on human 12q15.

Mdm2 is located on murine 10 D2.

PMIDs: 24309898, 29584707, 9840926, 28714398