Ubiquitin Rabbit Polyclonal Antibody

☑ Cell treatment (CT)

Western blot analysis of Ubiquitin on different lysates with Rabbit anti-Ubiquitin antibody (ER31212) at 1/500 dilution.

Lane 1: MCF-7 cell lysate
Lane 2: MCF-7 cell lysate treated with 50 µM MG132 for 1.5h

Lysates/proteins at 10 µg/Lane.

Predicted band size: 8 kDa

Exposure time: 2 minutes;
15% 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 (ER31212) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Western blot analysis of Ubiquitin on different cell lysates using anti-Ubiquitin antibody at 1/1,000 dilution.
Positive control:
Lane 1: 293T cell lysate
Lane 2: HepG2 cell lysate
Lane 3: Hela cell lysate
Lane 4: MCF-7 cell lysate

Immunocytochemistry analysis of NIH/3T3 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 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 Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunocytochemistry analysis of PC-12 cells labeling Ubiquitin with Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 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 Rabbit anti-Ubiquitin antibody (ER31212) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

ICC staining Ubiquitin in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

ICC staining Ubiquitin in HepG2 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

ICC staining Ubiquitin in MCF-7 cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.

Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ER31212) 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.

Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Ubiquitin antibody (ER31212) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ER31212) 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.