Anti-Sumo 1 antibody (ab11672)

Immunocytochemistry/ Immunofluorescence - Sumo 1 antibody (ab11672)

Immunofluorescence of 293T cells transfected with a vector that has Sumo1 fused to GFP and a Flag tag. The top panel is GFP fluorescence. The middle panel uses ab11672 at 1/300 with a rhodamine secondary antibody. The lower panel is a merge of the GFP fluorescence and ab11672 immunostaining. Staining was also seen in untransfected cells (although at a lower level).

Immunoprecipitation - Sumo 1 antibody (ab11672)

293T cells were transfected with a vector that has Sumo1 fused to GFP and a Flag tag. Cell lysates were used in IP with ab11672 (and a GFP antibody as a control). The resulting western blot was performed with a Flag antibody. As a control, cells were transfected with a a vector with Sumo2 fused to GFP and a Flag tag. ab11672 does not IP anything from this lysate.

Lane 1: Sumo1 fusion lysate - IP'd with GFP antibody

Lane 2: Sumo1 fusion lysate - no IP

Lane 3: Sumo1 fusion lysate - IP'd with ab11672

Lane 4: Sumo2 fusion lysate - IP'd with ab11672

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Sumo 1 antibody(ab11672)

Ab11672 staining human normal placenta. Staining is localized to the nucleus and nuclear membrane.
Left panel: with primary whole serum antibody at 1/400. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Immunocytochemistry/ Immunofluorescence - Sumo 1 antibody (ab11672)

ICC/IF image of ab11672 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 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 (ab11672, 1/1000 dilution) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) 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 1.43µM.