Key Product Details

Validated by

Biological Validation

Species Reactivity

Human, Mouse

Applications

ELISA, Immunoprecipitation, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all MDM2/HDM2 Antibodies »

Product Specifications

Immunogen

This affinity purified MDM2/HDM2 [p Ser185] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near aa 175-200 of mouse MDM2/HDM2/HDM2. (Uniprot: P23804)

Reactivity Notes

A BLAST analysis was used to suggest minimal cross reactivity with MDM2/HDM2 homologues from other sources.

Modification

p Ser185

Localization

Nuclear and cytoplasmic. Expressed predominantly in the nucleoplasm. Interaction with ARF (P14) results in the localisation of both proteins to the nucleus. The nucleolar localisation signals in both ARF and MDM2 may be necessary to allow efficient nucleo

Specificity

This affinity-purified antibody is directed against the phosphorylated form of mouse MDM2/HDM2 protein at the pS185 residue. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against Mouse MDM2/HDM2 pS185 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated mouse MDM2/HDM2 is minimal by ELISA and western blot. A BLAST analysis was used to suggest minimal cross reactivity with MDM2/HDM2 homologues from other sources.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This affinity-purified antibody is directed against the phosphorylated form of mouse MDM2 protein at the p Ser185 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for MDM2/HDM2 [p Ser185] Antibody

Western Blot: MDM2/HDM2 [p Ser185] Antibody [NB100-1683] - Shown in a 293T whole cell lysate. Cells were serum-starved for 24 hours prior to harvest. Approximately 20 ug of lysate was loaded per lane for SDS-PAGE. Untreated cells are shown in lane 1, whereas cells in lane 2 were treated with IGF-1 (100 ng/ml) for 20 min prior to harvest. Follow reaction of antibody with a 1:2000 dilution of HRP Goat-a-Rabbit IgG for visualization.
MDM2/HDM2 [p Ser185] Antibody

MDM2/HDM2 [p Ser185] Antibody

Affinity Purified Anti-MDM2 pS185 (Rabbit) is shown to detect a 102 kDa band (arrow) corresponding to phosphorylated mouse MDM2 present in a 293T whole cell lysate. Cells were serum-starved for 24 hours prior to harvest. Approximately 20 ug of lysate was loaded per lane for SDS-PAGE. Untreated cells are shown in lane 1, whereas cells in lane 2 were treated with IGF-1 (100 ng/ml) for 20 min prior to harvest. Follow reaction of antibody with a 1:2000 dilution of HRP Goat-a-Rabbit IgG for visualization.

Applications for MDM2/HDM2 [p Ser185] Antibody

Application
Recommended Usage

ELISA

1:3000-1:12000

Immunoprecipitation

1:100

Western Blot

1:500-1:2000
Application Notes
This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect bands approximately 102 kDa in size corresponding to phosphorylated MDM2 protein by western blotting in the appropriate cell lysate or extract.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: MDM2/HDM2

MDM2 is an E3 ubiquitin ligase that recognizes the N-terminal trans-activation domain (TAD) of the p53 transcription factor. MDM2 is a p53 responsive gene

Long Name

Double Minute 2 Protein

Alternate Names

HDM2

Entrez Gene IDs

4193 (Human)

Gene Symbol

MDM2

UniProt

P23804 (Mouse)

Additional MDM2/HDM2 Products

Product Documents for MDM2/HDM2 [p Ser185] Antibody

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for MDM2/HDM2 [p Ser185] Antibody

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.

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