Anti-SOX2 antibody [9-9-3]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody [9-9-3] (AB79351)

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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody [9-9-3] (AB79351)

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.

• ICC/IF

AbReview31574****

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody [9-9-3] (AB79351)

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.

This image is courtesy of an abreview by Joe Segal.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody [9-9-3] (AB79351)

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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody [9-9-3] (AB79351)

ab79351 staining SOX2 in mES cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab79351 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SOX2 antibody [9-9-3] (AB79351)

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 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.

• WB

Lab

Western blot - Anti-SOX2 antibody [9-9-3] (AB79351)

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

All lanes:

Western blot - Anti-SOX2 antibody [9-9-3] (ab79351) at 1/1000 dilution

Lane 1:

NCCIT (Human mediastinal epithelial pluripotent embryonal carcinoma cells) cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate adenocarcinoma cells) cell lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

Observed band size: 40 kDa

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