Anti-Ubiquitin antibody (ab14372)

Western blot - Ubiquitin antibody (ab14372)

Predicted band size : 8.5 kDa

ab14372 was tested against E.coli lysates expressing a ubiquitin GFP fusion protein and other UBL fusion proteins. All UBLs possess limited homology to Ubiquitin and to each other. Therefore it is important to know the degree of reactivity of each antibody against each UBL.

Panle A: shows total protein staining using Ponceau. Panel B: shows specific reaction with ubiquitin using a 1:1,000 and 1;2,000 dilution of ab14372 followed by an appropriate secondary antibody. The data indicate that ab14372 is highly specific and does not cross react with other UBLs.

Western blot - Ubiquitin antibody (ab14372)

Predicted band size : 8.5 kDa

ab14372 at dilution 1:200 (left lane) and 1:1000 (right lane) was tested by immunoblot against total cell extract from yeast. The blots shows strong reactivity of ab14372 with ubiquitinated proteins. Ab14372 was used at 1/500 dilution and incubated overnight at 4°C in 5% non-fat dry milk in TTBS.

Immunocytochemistry/ Immunofluorescence - Ubiquitin antibody (ab14372)

Central Panel: ab14372 staining Ubiquitin in human PMN cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 2% BSA for 1 hour. Samples were incubated with primary antibody 1/100 in blocking buffer for 4 hours at 37ºC. An Alexa Fluor^® 568-conjugated Goat polyclonal to rabbit IgG, dilution 1/250, was used as secondary antibody.
Left-hand panel: Nuclei counterstained with DAPI (blue).
Right-hand panel: Overlay

This image is courtesy of an anonymous Abreview

Flow Cytometry - Ubiquitin antibody (ab14372)

ab14372 staining Ubiquitin in Human platelet cells by Flow cytometry.
Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/200 dilution and incubated for 18 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.

Image courtesy of Dr Mahesh Shivananjappa by Abreview.