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MDM2 human protein exists in 11 isoforms UniProtKB - Q00987, the immunogen of ab38178 corresponds to atleast 10 isoforms i.e. 1,2,3,4,5,6 8, 9, 10 and 11; therefore the western blot results might show bands corresponding to these isoforms.
Our Abpromise guarantee covers the use of ab38618 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).|
|IHC-P||1/10 - 1/50.|
ICC/IF image of ab38618> stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab38618, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L)(ab150077) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling MDM2 with ab38618. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody, followed by DAB staining.
This image is courtesy of an anonymous AbreviewBlocked using 5% Milk for 30 minutes at Room temperature. Antibody Dilutent was5% milk in TBS-Tween-20 (0.05%). Primary incubation 3 hours at room temperature.
Immunofluorescence analysis of MDCK cells, staining MDM2 with ab38618.
Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 1% BSA in PBS) for 1 hour at 25°C. An Alexa Fluor® 488-conjugated goat anti-rabbit polyclonal IgG (1/500) (ab150077) was used as the secondary antibody.
ab38618 staining human cancer tissue by immunohistochemistry, formalin-fixed, paraffin-embedded tissue. The hepatocarcinoma tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining.
Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.
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