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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.
|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.