Thermo Fisher Scientific MDM2 Monoclonal Antibody (SMP14)

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:200

-

Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

SMP14

Immunogen

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Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Purification

Protein A

Storage buffer

tris with NP-40, BSA

Contains

<0.1% sodium azide

Storage conditions

4° C

Shipping conditions

Wet ice

Product Specific Information

A recommended positive control tissue for this product is Liposarcoma, however positive controls are not limited to this tissue type.

The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.

p53 is the most commonly mutated gene in human cancer identified to date. Expression of p53 leads to inhibition of cell growth by preventing progression of cells from G1 to S phase of the cell cycle. Most importantly, p53 functions to cause arrest of cells in the G1 phase of the cell cycle following any exposure of cells to DNA-damaging agents. The MDM2 (murine double minute-2) protein was initially identified as an oncogene in a murine transformation system. MDM2 functions to bind p53 and block p53-mediated transactivation of co-transfected reporter constructs. The MDM2 gene is amplified in a high percentage of human sarcomas that retain wide type p53 and tumor cells that overexpress MDM2 can tolerate high levels of p53 expression. These findings argue that MDM2 overexpression (gene amplification) represents at least one mechanism by which p53 function can be abrogated during tumorigenesis. MDM2 is useful in differentiating well-differentiated/dedifferentiated liposarcoma from other types of sarcomas.

Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.

A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.

Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.

Target Information

MDM2 is a ubiquitin ligase for p53 and plays a central role in regulation of the stability of p53. MDM2 is located on chromosome 12 on the q arm. Akt-mediated phosphorylation of MDM2 at Ser166 and Ser186 increases its interaction with p300, allowing MDM2-mediated ubiquitination and degradation of p53. Phosphorylation of MDM2 also blocks its binding to p19ARF, increasing the degradation of p53. MDM2 has also been shown to negatively regulate p53 function. MDM2 binds and inhibits transactivation role played by p53 and overexpression of MDM2 can result in the inactivation of p53 and decrease its tumor suppressor function. Another process by which MDM2 can inactivate p53 is by degrading p53 as the protein also possesses E3 ubiquitin ligase activity. Further, MDM2 plays important roles in apoptosis and cell cycle. MDM2 is over expressed in a wide range of human malignancies including soft tissue carcinomas and breast cancer. In addition to p53, MDM2 is involved in processes of cell cycle, apoptosis, and tumorigenesis through interactions with proteins that include retinoblastoma 1 and ribosomal protein L5. More than 40 different alternatively spliced transcript variants of MDM2 have been isolated from both tumor and normal tissues.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.