Human Total MDM2/HDM2 DuoSet IC ELISA

R&D Systems | Catalog # DYC1244-2

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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

312-20000 pg/mL

Sample Type

Cell Lysates

Reactivity

Human

Human Total MDM2/HDM2 DuoSet IC ELISA Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15- (96-well) plate pack sizes
  • Economical alternative to Western blot
View all MDM2/HDM2 ELISA Kits »

Product Summary for Human Total MDM2/HDM2 DuoSet IC ELISA

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure total MDM2 / HDM2 in cell lysates. An immobilized capture antibody specific for MDM2 / HDM2 binds both phosphorylated and unphosphorylated MDM2 / HDM2. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-HRP format.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

Cell lysates (100 µL)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human Total MDM2/HDM2 DuoSet IC ELISA

Standard Curve

Specificity of Human Total MDM2 Duoset IC ELISA is shown by Western blot.

Specificity of Human Total MDM2 Duoset IC ELISA is shown by Western blot.

Lysates prepared from LLnV treated U2OS human osteosarcoma cells were incubated in wells coated with Human Total MDM2 Capture Antibody. Unbound material was removed by washing and bound material was solubilized in SDS gel sample buffer. The same lysate and captured proteins were electrophoresed, transferred to a PVDF membrane, and immunoblotted with Human Total MDM2 Detection Antibody. The captured protein corresponds to MDM2 detected in the total cell lysates.
Amounts of MDM2, as quantified by the Human Total MDM2 Duoset IC ELISA, are consistent with the relative levels of MDM2 determined by qualitative Western blot analysis.

Amounts of MDM2, as quantified by the Human Total MDM2 Duoset IC ELISA, are consistent with the relative levels of MDM2 determined by qualitative Western blot analysis.

Lysates prepared from U2OS human osteosarcoma +/- LLnV cells were quantified with this DuoSet® IC. The same lysates were also immunoblotted (inset) with a monoclonal detection antibody. The DuoSet® IC results correlate well with the relative amounts of human MDM2 detected by Western Blot.

Kit Contents for Human Total MDM2/HDM2 DuoSet IC ELISA

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control
  • Streptavidin-HRP

Other Reagents Required


PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Preparation and Storage

Shipping

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

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: MDM2/HDM2

MDM2 is a key regulator of p53 tumor suppressor protein activity and stability. MDM2 binds to and inhibits the transactivation domain of p53. In addition, MDM2 controls p53 stability by functioning as its E3 ligase in ubiquitination and by shuttling p53 from the nucleus to the cytoplasm for subsequent degradation. The importance of the p53/MDM2 relationship is underscored by the existence of an autoregulatory feedback loop whereby activated p53 transcriptionally upregulates the expression of its own inhibitor, MDM2.

Long Name

Double Minute 2 Protein

Alternate Names

HDM2

Entrez Gene IDs

4193 (Human); 17246 (Mouse)

Gene Symbol

MDM2

Additional MDM2/HDM2 Products

Product Documents for Human Total MDM2/HDM2 DuoSet IC ELISA

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Human Total MDM2/HDM2 DuoSet IC ELISA

For research use only

Citations for Human Total MDM2/HDM2 DuoSet IC ELISA

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