Anti-SOX2 antibody [9-9-3]

7 product images

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  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 Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) 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.

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  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) (Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control 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.

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  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 1/10000 dilution

  Lane 1: NCCIT whole cell lysate at 10 µg

  Lane 2: PC-3 whole cell lysate at 10 µg

  Secondary

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

  Predicted band size: 34 kDa

  Observed band size: 34 kDa

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  This image is courtesy of a customer review by Joe Segal.

  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.

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  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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 AlexaFluor®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing showing positive staining on NCCIT cells. Counterstained with Anti-beta IV Tubulin antibody [EPR16775] ab179504 anti-Tubulin (Rabbit mAb, 1/1000), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at 1/1000 dilution.
  -ve control 2: Anti-beta IV Tubulin antibody [EPR16775] ab179504 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at 1/1000 dilution.

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  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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 AlexaFluor^®488 Goat anti-Mouse secondary antibody at 1/1000 dilution (green). Confocal image showing showing positive staining on F9 cells. Counterstained with Anti-beta IV Tubulin antibody [EPR16775] ab179504 anti-Tubulin (Rabbit mAb, 1/1000), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at 1/1000 dilution.
  -ve control 2: Anti-beta IV Tubulin antibody [EPR16775] ab179504 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at 1/1000 dilution.
  

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  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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) 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 Anti-beta IV Tubulin antibody [EPR16775] ab179504 anti-Tubulin (Rabbit mAb, 1/1000), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at 1/1000 dilution.
  -ve control 2: Anti-beta IV Tubulin antibody [EPR16775] ab179504 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at 1/1000 dilution.