Mdm4 / HdmX-Trap

Details for Product No. ABIN1889486, Supplier: Log in to see
Antigen
  • mdmx
  • HDMX
  • MDMX
  • MRP1
  • wu:fa09h09
  • wu:fi33d10
  • 4933417N07Rik
  • AA414968
  • AL023055
  • AU018793
  • AU021806
  • C85810
  • Mdmx
  • Mdm4 p53 binding protein homolog
  • Mdm4 p53 binding protein homolog (mouse)
  • transformed 3T3 cell double minute 4 homolog (mouse)
  • transformed mouse 3T3 cell double minute 4
  • mdm4
  • MDM4
  • Mdm4
Alternatives
Human MDM4-binding Protein ELISA Kit
Epitope
AA 1-129
Reactivity
Human
5
3
3
Host
Camelidae
Application
Affinity Measurement (AM), Chromatin Immunoprecipitation (ChIP), Enzyme Activity Assay (EAA), Immunoprecipitation (IP), Mass Spectrometry (MS), Protein Complex Immunoprecipitation (Co-IP), Pull-Down Assay (Pull-Down), Purification (Purif)
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Purpose The Mdm4/ HdmX-Trap is a high quality Mdm4/ HdmX-binding protein for biochemical analysis of Mdm4/ HdmX and interacting partners.
Sample Type Cell Extracts
Specificity Species-Reactivity: tested on human, mouse, hamster
Characteristics Mdm4/ HdmX-Trap is excellent for fast and efficient one-step isolation of Mdm4/ HdmX and its interacting factors from cellular extract. Isolated Mdm4/ HdmX protein may be used further for immunoblot analysis or mass spectrometry.
Mdm4/ HdmX-Trap utilizes small recombinant alpaca antibody fragments covalently coupled to the surface of agarose beads.
Purification Purifed protein
Components Mdm4/ HdmX-Trap uncoupled
Material not included Lysis buffer (CoIP), 10x RIPA buffer, Dilution buffer, Wash buffer, Elution buffer
Alternative Name Mdm4 (MDM4 ELISA Kit Abstract)
Background Double minute 4 protein (Mdm4/ HdmX/ MdmX) plays a role in cell cylce control and apoptosis by inhibition of p53 and p73.
Comment

Use this Mdm4/HdmX-Trap to couple it to your own immobilizing matrix.
Note that binding of Mdm4/ HdmX-Trap interferes with binding of p53 to Mdm4/ HdmX.

Assay Time 1.5 h
Protocol
  • Robust and versatile tool for biochemical analyses of Mdm4/ HdmX proteins
  • Short incubation times (5 - 30 min)
  • Quantitative isolation of fusion proteins and transiently bound factors from cell extracts or organelles
  • Low unspecific binding
  • No contaminating heavy and light chains of conventional antibodies
  • Applicable in Chromatin Immunoprecipitation (ChIP)
Sample Collection Harvest cells:
For one immunoprecipitation reaction the use of 10^6 - 10^7 mammalian cells (approx. one 10 cm dish) is recommended. To harvest adherent cells, aspirate growth medium, add 1 ml ice-cold PBS to cells and scrape cells from dish. Transfer cells to a pre-cooled tube, spin at 500 g for 3 min at +4°C and discard supernatant. Wash cell pellet twice with ice-cold PBS, gently resuspending the cells.

Lyse cells
  • 1. Resuspend cell pellet in 200 µL ice-cold lysis buffer by pipetting or using a syringe.
    note: Supplement lysis buffer with protease inhibitors and 1 mM PMSF (not included). optional for nuclear/chromatin proteins: Use RIPA buffer supplemented with 1 mg/mL DNase, 2.5 mM MgCl2, protease inhibitors and 1 mM PMSF (not included).
  • 2. Place the tube on ice for 30 min with extensively pipetting every 10 min.
  • 3. Centrifuge cell lysate at 20.000x g for 10 min at +4°C. Transfer lysate to a pre-cooled tube. Add 300 µl dilution buffer to lysate. Discard pellet.
    note: At this point cell lysate may be put at -80°C for long-term storage. optional: Add 1 mM PMSF and protease inhibitors (not included) to dilution buffer

We recommend that during incubation with the beads the final concentration of detergents does not exceed 0.2% to avoid unspecific binding to the matrix. If required, use more dilution buffer to dilute the supernatant accordingly.
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer 1 x PBS, 0.01 % Sodium azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Do not freeze.
Storage 4 °C
Expiry Date 12 months
Supplier Images
Immunoprecipitation (IP) image for Mdm4 / HdmX-Trap (ABIN1889486) Pulldown of Mdm4/HdmX using the Mdm4/HdmX-Trap:Immunoprecipitation (IP) of Mdm4/HdmX ...