MDM2/HDM2 Antibody (A10E11-R)

Western Blot: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584]

Western Blot: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Western blot analysis of MDM2/HDM2 on different lysates with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) at 1/1,000 dilution.

Lane 1: U-2 OS cell lysate
Lane 2: U-2 OS treated with 10uM NUTLIN 3A for 24 hours cell lysate

Lysates/proteins at 15 ug/Lane.

Predicted band size: 55 kDa
Observed band size: 90 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32584) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemistry: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584]

Immunohistochemistry: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Immunohistochemical analysis of paraffin-embedded human embryonal carcinoma tissue with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32584) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584]

Immunocytochemistry/ Immunofluorescence: MDM2/HDM2 Antibody (A10E11-R) [NBP3-32584] - Immunocytochemistry analysis of U-2 OS cells treated with or without 10μM NUTLIN 3A for 24 hours labeling MDM2/HDM2 with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-MDM2/HDM2 antibody (NBP3-32584) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.