ab134953 staining Ubiquitin in Mouse embryo tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer, pH6. Samples were incubated with primary antibody (1/100 in 1% FBS/BSA) for 16 hours at 4°C. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.See Abreview
Overlay histogram showing HepG2 cells stained with ab134953 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab134953, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
![All lanes : Anti-Ubiquitin antibody [EPR8830] (ab134953) at 1/1000 dilutionLane 1 : 293T cell lysateLane 2 : HepG2 cell lysateLane 3 : HeLa cell lysateLane 4 : JAR cell lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti rabbit at 1/2000 dilution](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/17519_Ubiquitin-Primary-antibodies-ab134953-4.jpg)
All lanes : Anti-Ubiquitin antibody [EPR8830] (ab134953) at 1/1000 dilutionLane 1 : 293T cell lysateLane 2 : HepG2 cell lysateLane 3 : HeLa cell lysateLane 4 : JAR cell lysateLysates/proteins at 10 µg per lane.SecondaryHRP labelled goat anti rabbit at 1/2000 dilution
Immunofluorescent staining of JAR cells labelling Ubiquitin using ab134953 at 1/250 dilution
Immunohistochemical analysis of paraffin embedded Human breast carcinoma tissue labelling Ubiquitin with ab134953 at 1/250 dilution