MDM2 ELISA Kit

Catalog No. ABIN8070011

Human MDM2 ELISA Kit, Colorimetric assay for quantification of Human MDM2.

Quick Overview for MDM2 ELISA Kit (ABIN8070011)

Target: MDM2 (Mdm2, p53 E3 Ubiquitin Protein Ligase Homolog (Mouse) (MDM2))

Binding Specificity: SUMOylated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details MDM2 ELISA Kit

Purpose: Measures sumoylated MDM2 and total MDM2 in human cell and tissue lysates, enabling precise assessment of MDM2 sumoylation by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-MDM2 Antibody
• Anti-SUMOylation Detection Antibody
• Anti-MDM2 Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Target Details for MDM2

Target: MDM2 (Mdm2, p53 E3 Ubiquitin Protein Ligase Homolog (Mouse) (MDM2))

Alternative Name: MDM2

Background:

Gene Symbols: MDM2

Background: The SUMOylated-MDM2 and Total-MDM2 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human sumoylated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan MDM2 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and MDM2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-sumoylation antibody is used to detect sumoylated MDM2 or biotinylated anti-MDM2 antibody is used to detect total MDM2 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of sumoylated MDM2 or total MDM2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): E3 ubiquitin-protein ligase Mdm2, Double minute 2 protein (Hdm2), Oncoprotein Mdm2, RING-type E3 ubiquitin transferase Mdm2, p53-binding protein Mdm2

Gene ID: 4193

UniProt: Q00987

Pathways: p53 Signaling, PI3K-Akt Signaling, Cell Division Cycle, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Autophagy, Ubiquitin Proteasome Pathway