Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonal carcinoma) cells labeling SOX2 with ab79351 at 1/200 dilution, followed by ab150113 AlexaFluor®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing showing positive staining on NCCIT cells. Counterstained with ab179504 anti-Tubulin (Rabbit mAb, 1/1000), ab150080 AlexaFluor®594 Goat anti-Rabbit secondary at a 1/1000 dilution. Nuclear stained with DAPI.
The negative controls are as follows:
-ve control 1: ab79351 at 1/200 dilution, followed by ab150080 at 1/1000 dilution.
-ve control 2: ab179504 at 1/1000 dilution, followed by ab150113 at 1/1000 dilution.

All lanes : Anti-SOX2 antibody [9-9-3] (ab79351) at 1/10000 dilution

Lane 1 : NCCIT whole cell lysate
Lane 2 : PC-3 whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST

Overlay histogram showing F9 cells stained with ab79351 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab79351, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in 80% methanol (5 min) fixed F9 cells used under the same conditions.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma) cells labeling SOX2 with ab79351 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing showing positive staining on F9 cells. Counterstained with ab179504 anti-Tubulin (Rabbit mAb, 1/1000), ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at a 1/1000 dilution. Nuclear stained with DAPI.
The negative controls are as follows:
-ve control 1: ab79351 at 1/200 dilution, followed by ab150080 at 1/1000 dilution.
-ve control 2: ab179504 at 1/1000 dilution, followed by ab150113 at 1/1000 dilution.

 

 

ICC/IF image of ab79351 stained D3 mouse embryonic stem cells. The cells were fixed in 4% Paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes at RT, before incubation with ab79351 at a 1/100 dilution for 2 hours at RT. The secondary used was an Alexa Fluor 488 conjugated goat anti-mouse polyclonal, used at 1/200 dilution.

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Immunofluorescent analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) (Negative control) cells labeling SOX2 with ab79351 at 1/200 dilution, followed by ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing showing no staining on NIH/3T3 cells. Counterstained with ab179504 anti-Tubulin (Rabbit mAb, 1/1000), ab150080 AlexaFluor^®594 Goat anti-Rabbit secondary at a 1/1000 dilution. Nuclear stained with DAPI.
The negative controls are as follows:
-ve control 1: ab79351 at 1/200 dilution, followed by ab150080 at 1/1000 dilution.
-ve control 2: ab179504 at 1/1000 dilution, followed by ab150113 at 1/1000 dilution.