Anti-Ubiquitin antibody [P4D1]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ubiquitin antibody [P4D1] (AB303664)

Flow cytometric (Intracellular) overlay histogram showing 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Ubiquitin with ab303664 at 1/800 dilution (0.1μg) (red line). Isotype control antibody (black line) was a mouse monoclonal IgG used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [P4D1] (AB303664)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Ubiquitin with ab303664 at 1/20000 (0.044 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Nuclear staining on human colon cancer. The section was incubated with ab303664 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody Leica DS9800 (Bond™ Polymer Refine Detection kit). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [P4D1] (AB303664)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Ubiquitin with ab303664 at 1/20000 (0.044 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Nuclear staining on rat cerebrum. The section was incubated with ab303664 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody Leica DS9800 (Bond™ Polymer Refine Detection kit). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [P4D1] (AB303664)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Ubiquitin with ab303664 at 1/20000 (0.044 μg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection kit). Nuclear staining on mouse cerebrum. The section was incubated with ab303664 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Primary diluent was used instead of primary antibody, followed by a ready to use secondary antibody Leica DS9800 (Bond™ Polymer Refine Detection kit). Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ubiquitin antibody [P4D1] (AB303664)

Flow cytometric (Intracellular) overlay histogram showing 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Ubiquitin with ab303664 at 1/800 dilution (0.1μg) (red line). Isotype control antibody (black line) was a mouse monoclonal IgG used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

• WB

Supplier Data

Western blot - Anti-Ubiquitin antibody [P4D1] (AB303664)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 26 seconds

All lanes:

Western blot - Anti-Ubiquitin antibody [P4D1] (ab303664) at 1/1000 dilution

Lane 1:

Untreated 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

293T treated with 10μM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 4 hours, whole cell lysate at 20 µg

Lane 3:

Untreated HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HeLa treated with 10μM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 6 hours, whole cell lysate at 20 µg

Lane 5:

Untreated MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

MCF7 treated with 10μM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 2 hours, whole cell lysate at 20 µg

Lane 7:

Untreated NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 8:

NIH/3T3 treated with 20μM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 5 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 8 kDa,200 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Ubiquitin antibody [P4D1] (AB303664)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 3 seconds

All lanes:

Western blot - Anti-Ubiquitin antibody [P4D1] (ab303664) at 1/1000 dilution

Lane 1:

Untreated PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Lane 2:

PC-12 treated with 10μM MG-132 (<a href='/ko/products/biochemicals/mg-132-proteasome-inhibitor-ab141003'>ab141003</a>) for 4 hours, whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 8 kDa,200 kDa

false

Exposure time: 3s