Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 09, 2021

Anti-MDM2 antibody [EPR22256-98] (ab259265)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22256-98] to MDM2
Suitable for: ICC/IF, IP, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-MDM2 antibody [EPR22256-98]
See all MDM2 primary antibodies
Description

Rabbit monoclonal [EPR22256-98] to MDM2
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: RAW 264.7 treated with 10uM Nutlin-3a for 24 hours whole, 2.4G2 treated with 10uM Nutlin-3a for 24 hours whole lysates. ICC/IF: HepG2 cells treated with 10uM Nutlin-3a for 24 hours. IP: HepG2 treated with 10uM Nutlin-3a for 24 hours whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22256-98
Isotype

IgG
Research areas

Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265)

All lanes : Anti-MDM2 antibody [EPR22256-98] (ab259265) at 1/1000 dilution

Lane 1 : Untreated RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 2 : RAW 264.7 treated with 10uM Nutlin-3a for 24 hours, whole cell lysate
Lane 3 : Untreated 2.4G2 (rat B cell lymphoma B lymphocyte), whole cell lysate
Lane 4 : 2.4G2 treated with 10uM Nutlin-3a for 24 hours, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 55 kDa
Observed band size: 60,90 kDa
why is the actual band size different from the predicted?

Blocking and Dilution Buffer and concentration: 5% NFDM/TBST.

MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53-MDM2 interaction and induces p53- dependent apoptosis and autophagy.

The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).

Exposure time: 15 seconds.
Western blot - Anti-MDM2 antibody [EPR22256-98] (ab259265)

All lanes : Anti-MDM2 antibody [EPR22256-98] (ab259265) at 1/1000 dilution

Lane 1 : Untreated HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate
Lane 2 : Treated for 24 hours HepG2 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 55 kDa
Observed band size: 60,90 kDa why is the actual band size different from the predicted?

Blocking and Dilution Buffer and concentration: 5% NFDM/TBST.

MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53-MDM2 interaction and induces p53- dependent apoptosis and autophagy.

The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).

Exposure time: 10 seconds.
Immunoprecipitation - Anti-MDM2 antibody [EPR22256-98] (ab259265)

MDM2 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) treated with 10uM Nutlin-3a for 24 hours whole cell lysate 10ug with ab259265 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259265 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) treated with 10uM Nutlin-3a for 24 hours whole cell lysate 10ug

Lane 2: ab259265 IP in HepG2 treated with 10uM Nutlin-3a for 24 hours whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259265 in HepG2 treated with 10uM Nutlin-3a for 24 hours whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 seconds.

MDM2 can be cleaved into a 60-kDa fragment after Nutlin 3a treatment as Nutlin 3a disrupts p53–MDM2 interaction and induces p53- dependent apoptosis and autophagy.

The molecular weight observed is consistent with what has been described in the literature (PMID:19638413, 10329737).
Immunocytochemistry/ Immunofluorescence - Anti-MDM2 antibody [EPR22256-98] (ab259265)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling MDM2 with ab259265 at 1/100 (5 ug/ml) dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing nuclear staining in HepG2 cells treated with Nutlin-3a (10 uM) for 24 hours is observed. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab259265 anti- MDM2 ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 (2 ug/ml) dilution.
Anti-MDM2 antibody [EPR22256-98] (ab259265)

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