Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue sections labeling Pumilio 1 with Purified ab92545 at 1:100 dilution (1.18 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling Pumilio 1 with purified ab92545 at 1:20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Pumilio 1 with purified ab92545 at 1:500 dilution (0.2μg/ml). Cells were fixed in 100% Methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

All lanes : Anti-Pumilio 1 antibody [EPR3795] (ab92545) at 1/1000 dilution (unpurified)

Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 126 kDa

Immunohistochemical analysis of Pumilio 1 in paraffin embedded Human fetal kidney tissue using an unpurified ab92545 at a 1/100 dilution.

All lanes : Anti-Pumilio 1 antibody [EPR3795] (ab92545) at 1/1000 dilution (purified)

Lane 1 : Mouse cerebellum lysates
Lane 2 : Rat brain lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 126 kDa

Blocking and diluting buffer: 5% NFDM/TBST 

All lanes : Anti-Pumilio 1 antibody [EPR3795] (ab92545) at 1/5000 dilution (purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 126 kDa

Blocking and diluting buffer: 5% NFDM/TBST

Immunofluorescent staining of Pumilio 1 in HeLa cells using an unpurified ab92545 at a 1/100 dilution.

Overlay histogram showing HEK293 cells stained with an unpurified ab92545 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab92545, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1 µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.