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Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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- Product number
- MA1-24755 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-MDM2 Monoclonal Antibody (HDM2-323)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- HDM2-323
- Vial size
- 100 µL
- Concentration
- 2 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot of MDM2 in whole extracts of HEK-293 cells overexpressing human (lane 1) or mouse (lane 2) MDM2 using a MDM2 monoclonal antibody (Product # MA1-24755) at a dilution of 2 µg/mL followed by detection using a Goat anti-mouse IgG-Peroxidase and chemiluminescent substrate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of MDM2 was achieved by transfecting U-2 OS with MDM2 specific siRNAs (Silencer® select Product # S8628, S8629). Western blot analysis (Fig. a) was performed using Whole cell extracts from the MDM2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with MDM2 Monoclonal Antibody (HDM2-323) (Product # MA1-24755, 1 ug/ml ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to MDM2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MDM2 Monoclonal Antibody (HDM2-323) (Product # MA1-24755) and a 75kDa band corresponding to N-terminally truncated MDM2 isoform was observed across cell lines tested. A non specific band around 150kD was also observed. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), HEK-293 (Lane 2) and MDA-MB-231 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
