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Primary Antibodies  >  Cell Cycle Protein Antibodies
 

MDM2 phospho S185 Antibody

Rabbit Polyclonal
Code

600-401-423
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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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.  Approxi-mately 20 µg 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.
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Synonyms: E3 ubiquitin-protein ligase Mdm2 EC=6.3.2. p53-binding protein Mdm2 Oncoprotein Mdm2 Double minute 2 protein

MDM2 phospho S185 Antibody Properties

Description
Anti-MDM2 pS185 (RABBIT) Antibody - 600-401-423
Host
Rabbit
Target Species
Mouse
Known Cross Reactivity
human, mouse, rat
Clonality
Polyclonal
Application
ELISA : 1:3,000 - 1:12,000
Western Blot : 1:500 - 1:2,000
ImmunoPrecipitation: 1:100
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.12 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
MDM2
Download CofA
Lot#  

MDM2 phospho S185 Antibody Description

Background
MDM2 is a nuclear phosphoprotein with an apparent molecular mass of 90 kD that forms a complex with the p53 tumor suppressor protein. Human MDM2 was identified as a homologous product of the 'murine double minute 2' gene (mdm2). The MDM2 gene enhances the tumorigenic potential of cells when it is overexpressed and encodes a putative transcription factor. Forming a tight complex with the p53 gene, the MDM2 oncogene can inhibit p53-mediated transactivation. MDM2 binds to p53 and amplification of MDM2 in sarcomas leads to escape from p53-regulated growth control. This mechanism of tumorigenesis parallels that for virus-induced tumors in which viral oncogene products bind to and functionally inactivate p53. Overexpression of the MDM2 oncogene was found in leukemias. Inactivation of tumor suppressor genes leads to deregulated cell proliferation and is a key factor in human tumorigenesis. MDM2 interacts physically and functionally with the retinoblastoma (RB) protein and can inhibit its growth regulatory capacity. Both RB and p53 can be subjected to negative regulation by the product of a single cellular protooncogene. The interference of binding to p53 prevents the interaction of MDM2 and its regulation of the transcriptional activity of p53 in vivo. Direct association of p53 with the cellular protein MDM2 results in ubiquitination and subsequent degradation of p53. MDM2-p53 complexes were preferentially found in S/G2M phases of the cell cycle. The MDM2 gene is alternatively spliced, producing 5 additional splice variant transcripts from the full length MDM2 gene. Four out of five of these alternatively spliced forms (MDM2a-MDMd) are missing substantial portions of the p53 binding domain and retain the acidic domain and the zinc-finger domains. The fifth and smallest transcript (MDM2e) retains the largest spliced region encoding the p53 binding domain; however, it lacks the nuclear localization signal, the acidic domain and zinc-finger domains. The alternatively spliced transcripts tend to be expressed in tumorigenic tissue, whereas the full-length MDM2 transcript is expressed in normal tissue. MDM2 is found in the nucleus and cytoplasm, however, it is expressed predominantly in the nucleoplasm. Interaction with ARF (P14) results in the localization of both proteins to the nucleus. The nucleolar localization signals in both ARF and MDM2 may be necessary to allow efficient nucleolar localization of both proteins.
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to aa 177-198 of mouse MDM2.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening.  Aliquot contents and freeze at -20° C 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 4° C as an undiluted liquid.  Dilute only prior to immediate use.
Application Note
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.
Purity/Specifity
This affinity-purified antibody is directed against the phosphorylated form of mouse MDM2 protein at the pS185 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.  Reactivity occurs against Mouse MDM2 pS185 protein and the antibody is specific for the phosphorylated form of the protein.   Reactivity with non-phosphorylated mouse MDM2 is minimal by ELISA and western blot.  A BLAST analysis was used to suggest minimal cross reactivity with MDM2 homologues from other sources.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
Pospisilova S, Siligan C, Ban J, Jug G, Kovar H. (2004) Constitutive and DNA Damage Inducible Activation of pig3 and MDM2 Genes by Tumor-Derived p53 Mutant C277Y. Mol Cancer Res. 2(5):296-304. Aslanian A, Iaquinta PJ, Verona R, Lees JA. (2004) Repression of the Arf tumor suppressor by E2F3 is required for normal cell cycle kinetics. Genes Dev. 2004 Jun 2 [Epub ahead of print]. Stros M, Muselikova-Polanska E, Pospisilova S, Strauss F. (2004) High-Affinity Binding of Tumor-Suppressor Protein p53 and HMGB1 to Hemicatenated DNA Loops. Biochemistry. 43(22):7215-7225.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IP, ChIP
Reacts With
human
Catalog Number
200-301-174

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
yeast
Catalog Number
200-301-428

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, bovine, dog, macaque, chimpanzee, rat
Catalog Number
600-401-859

Product Type
Primary Antibodies;
Applications
ELISA, WB, IP
Reacts With
mouse
Catalog Number
600-401-927

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry