Immunofluorescence analysis of MDM2 was done on 70% confluent log phase U2OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ MDM2 Recombinant Rabbit Monoclonal Antibody (700555) at 2 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant protein corresponding to amino acids 215-422 of human MDM2.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-3 ug/ml|
|Immunofluorescence (IF)||1-3 ug/ml|
|Western Blot (WB)||0.1-1 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
70-0555 has been successfully used in immunofluorescence and western blot. In western blot applications, 70-0555 recognizes the ~55kD and ~75kD isoforms of MDM2.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
MDM2 also known as murine double minute 2, is found to interact with p53; a tumor suppressor protein, and has been shown to negatively regulate its function. MDM2 binds and inhibits transactivation role played by p53 and overexpression of MDM2 can result in the inactivation of p53 and in turn decrease its tumor suppressor function. Another process by which MDM2 can inactivate p53 is by degrading p53 as the protein also possesses E3 ubiquitin ligase activity. MDM2 also plays various important roles in apoptosis and cell cycle. MDM2 is located on chromosome 12 on the q arm.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.