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- Antigen structure
- References [2]
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- Validations
- Immunocytochemistry [1]
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- Product number
- PA5-13008 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-MDM2 (Ser395) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 0.35 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references Characterization of SMG7 14-3-3-like domain reveals phosphoserine binding-independent regulation of p53 and UPF1.
SMG7 is a critical regulator of p53 stability and function in DNA damage stress response.
Cowen LE, Luo H, Tang Y
Scientific reports 2019 Sep 11;9(1):13097
Scientific reports 2019 Sep 11;9(1):13097
SMG7 is a critical regulator of p53 stability and function in DNA damage stress response.
Luo H, Cowen L, Yu G, Jiang W, Tang Y
Cell discovery 2016;2:15042
Cell discovery 2016;2:15042
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Phospho-MDM2 (Ser395) was performed using 70% confluent log phase HeLa cells treated with 0.5uM Doxorubicin for 24 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-MDM2 (Ser395) Polyclonal Antibody (Product # PA5-13008) at 1:100 dilution in 0.1% BSA, incubated overnight at 4 degree Celsius and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents the untreated cells with negligible expression of Phospho-MDM2 (Ser395). Panel f shows control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Dot blot analysis of Phospho-MDM2 pSer395 polyclonal antibody (Product # PA5-13008) on nitrocellulose membrane. 50 ng of Phospho-peptide or Non Phospho-peptide per dot were adsorbed. Antibody working concentrations are 0.5 µg per mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 Disruption of stressed-induced SMG7 binding to p53 does not significantly affect p53 stabilization, activation or DNA damage response. ( a ) Western blot analysis of cell extracts from control and irradiated (6 hours) cells of various SMG7 genotypes. ( b ) alpha-MDM2 immunoprecipitates from control or irradiated cells were assayed for Ser395 phosphorylation. ( c ) Cell extracts from control or Doxo-treated (0.4 muM) cells were analyzed by western blot using antibodies as indicated. ( d , e ) Control or Doxo-treated (0.4 muM, 16 hours) cells were fed with 10 muM CldU for 1 hour and then processed for CldU (green) and DAPI (blue) staining, followed by fluorescence microscopy. Representative images were shown in ( d ), and quantification of CldU-positive cells from five independent experiments in ( e ). (*p < 0.05, **p < 0.005; error bars represent SEM). ( f ) Western blot analysis of cell extracts from control or CPT-treated (1 µM) cells using antibodies as indicated. ( g ) A working model for SMG7 regulation of p53 stability upon DNA damage.
