ab7780 staining Ubiquitin in Panc-1 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in formaldehyde, blocked with 1% serum for 1 hour at 22°C and then incubated with ab7780 at a 1/2000 dilution for 1 hour at 22°C. The secondary was used at a 1/200 dilution.Green - Golgi residental proteins C2GnT-M.Red -Ubiquitin.Blue - nucleus staining by DAPI.

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ab7780 staining ubiquitin in cells transfected with FGAMS-EGFP by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with 3.7% methanol-free formaldehyde at 37°C for 15-20 minutes, blocked with 5% goat serum in PBS-T buffer for 30-60 minutes at room temperature. An Alexa Fluor® 594-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody

ICC/IF image of ab7780 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7780, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.

ab7780 (1/100) staining Ubiquitin in paraffin-embedded brain sections of a patient with Alzheimer's disease.

ab7780 staining ubiquitin in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/90) for 16 hours at 4°C. A HRP-conjugated Goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.

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ab7780 at a dilution of 1/1000, staining Ubiquitin in neurons (Alexa 488 secondary at 1/2000) on 30µm coronal rat brain (ab29475) tissue sections in free floating IHC (see protocol link for detailed description). Image shows neuronal staining observed with [A] 20x objective and [B] 40x objective.

NB: No labeling observed following omission of primary antibody.

Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).