Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (AB247278)

This data was developed using ab33893, the same antibody clone in a different buffer formulation.

ab33893 at a 1 : 250 dilution staining Ubiquitin in human prostate adenocarcinoma using Immunohistochemistry, Paraffin Embedded Tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (AB247278)

This data was developed using ab33893, the same antibody clone in a different buffer formulation.

IHC image of Ubiquitin staining in mouse Alzheimer brain formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33893, 1in500, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Lab

Western blot - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (AB247278)

This data was developed using ab33893, the same antibody clone in a different buffer formulation.

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. ab33893 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-Ubiquitin antibody [EP296Y] (<a href='/en-us/products/primary-antibodies/ubiquitin-antibody-ep296y-ab33893'>ab33893</a>) at 1/1000 dilution

Lane 1:

MCF-7 Whole cell lysate at 20 µg

Lane 2:

MCF-7 Whole cell lysate + (50 uM 90 min) at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Predicted band size: 26 kDa

false

• WB

Unknown

Western blot - Anti-Ubiquitin antibody [EP296Y] - BSA and Azide free (AB247278)

This data was developed using ab33893, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Ubiquitin antibody [EP296Y] (<a href='/en-us/products/primary-antibodies/ubiquitin-antibody-ep296y-ab33893'>ab33893</a>) at 1/2000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

Predicted band size: 26 kDa

Observed band size: 8.5 kDa

false