Confocal image showing nuclear staining on F9 cells

Ab92494 staining SOX2 in the F9 (mouse embryonal carcinoma) cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA, permeabilized with 0.1% Triton-X. Samples were incubated with primary antibody (1/200). An Alexa Fluor^® 488-conjugated Goat anti-Rabbit IgG, Ab150077 (1/1000) was used as the secondary antibody. Counterstained with Ab7291 anti-Tubulin (1/1000), Ab150120 AlexaFluor^®594 Goat anti-Mouse secondary (1/1000). DAPI was used as a nuclear counter stain.

Negative control 1 Ab92494 was used as the primary antibody at 1/200 and Ab150120 was used as the secondary at 1/1000.

Negative control 2 Ab7291was used as the primary antibody at 1/1000 and Ab150077 was used as the secondary at 1/1000.

ab92494 staining SOX2 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with , 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 ab92494 at 1/100 dilution and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

Transient Wnt and FGF signalling induce dual fated mouse neuromesodermal progenitors.

Immunostaining of cells treated with FGF/Wnt revealed the coexpression of Brachyury with Sox2 (NMPs). In the absence of Wnt, NPCs express Sox2 but the expression of Brachyury is only evident in a very small proportion of cells.

SOX2 immunostaining in sagittal maxillary incisor sections from E12 (A-D), E13 (E-H), E14 (I-L), and E15 (M-P) embryos.

At E13, strong SOX2 staining was seen in the lingual region of the epithelial dental lamina in all mice (E, G & H) except for the Usag-1^+/+/Runx2^-/- mice, in which SOX2 was found throughout the dental lamina (F). At E15, strong SOX2 staining was seen in the additional lingual bud in the Usag-1^+/+/Runx2^-/- mice (N).

All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution

Lane 1 : NCCIT (human pluripotent embryonic carcinoma cell line) whole cell lysate
Lane 2 : PC-3 (human prostate adenocarcinoma cell line) whole cell lysate
Lane 3 : SK-OV-3 (human ovarian cancer cell line) whole cell lysate
Lane 4 : U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate
Lane 5 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate
Lane 6 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 7 : Human breast cancer tissue lysate
Lane 8 : Human glioma lysate

Lysates/proteins at 10 µg per lane.

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

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


Exposure time: 3 minutes

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

Confocal image showing nuclear staining on NCCIT cells

Ab92494 staining SOX2 in NCCIT cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA, permeabilized with 0.1% Triton-X. Samples were incubated with primary antibody (1/200). An Alexa Fluor^® 488-conjugated Goat anti-Rabbit IgG, Ab150077 (1/1000) was used as the secondary antibody. Counterstained with Ab7291 anti-Tubulin (1/1000), Ab150120 AlexaFluor^®594 Goat anti-Mouse secondary (1/1000). DAPI was used as a nuclear counter stain.

Negative control 1 Ab92494 was used as the primary antibody at 1/200 and Ab150120 was used as the secondary at 1/1000.

Negative control 2 Ab7291was used as the primary antibody at 1/1000 and Ab150077 was used as the secondary at 1/1000.

All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution

Lane 1 : F9 (mouse embryonic testicular cancer cell line) whole cell lysate
Lane 2 : 4T1 (mouse mammary gland carcinoma cell line) whole cell lysate
Lane 3 : Mouse hippocampus lysate
Lane 4 : C6 (rat glial tumor cell line) whole cell lysate
Lane 5 : Rat hippocampus lysate
Lane 6 : Rat spinal cord lysate

Lysates/proteins at 10 µg per lane.

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

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


Exposure time: 3 minutes

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

Confocal image showing negative staining on NIH/3T3 cells.

Ab92494 staining SOX2 in the NIH/3T3 (mouse embryonic fibroblast cell line) (negative control) cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA, permeabilized with 0.1% Triton-X. Samples were incubated with primary antibody (1/200). An Alexa Fluor^® 488-conjugated Goat anti-Rabbit IgG, ab150077 (1/1000) was used as the secondary antibody. Counterstained with ab7291 anti-Tubulin (1/1000), Ab150120 Alexa Fluor^® 594 Goat anti-Mouse secondary (1/1000). DAPI was used as a nuclear counter stain.

Negative control 1: ab92494 was used as the primary antibody at 1/200 and ab150120 was used as the secondary at 1/1000.

Negative control 2: ab7291was used as the primary antibody at 1/1000 and ab150077 was used as the secondary at 1/1000.

Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution (unpurified) + NCCIT (human pluripotent embryonic carcinoma cell line) cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1500 dilution (purified) + F9 (mouse embryonic testicular cancer cell line) cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/5000 dilution (unpurified)

Lane 1 : NCCIT (human pluripotent embryonic carcinoma cell line) cell lysate
Lane 2 : MCF-7 (human breast adenocarcinoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 34 kDa

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling SOX2 with unpurified ab92494 at 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling SOX2 with purified ab92494 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling SOX2 with unpurified ab92494.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal stomach tissue labelling SOX2 with unpurified ab92494.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal lung tissue labelling SOX2 with unpurified ab92494.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human embryonal carcinoma tissue labelling SOX2 with unpurified ab92494.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human lung tissue. Unpurified ab92494 shows negative staining.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

Negative control: Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of negative human seminoma tissue using unpurified ab92494.

Heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 was performed before commencing with IHC staining protocol.

IHC - Wholemount analysis of Leucoraja erinacea embryo labelling SOX2 with unpurified ab92494 at 1/200. The sample was incubated with the primary antibody for 48 hours at 4°C in 10% fetal calf serum in PBT. Detection: DAB.

See Abreview

IHC - Wholemount analysis of mouse blastocyst labelling SOX2 (pink) with unpurified ab92494 at 1/200. The sample was incubated with the primary antibody for 48 hours at 4°C. Nuclei stained with DAPI (grey).

See Abreview

Standard Curve for SOX2 (Analyte: SOX2 protein (Human) (ab79950)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [57CT23.3.4] to SOX2 (ab75485) at 0.2µg/ml and Detector Antibody Rabbit monoclonal [EPR3131] to SOX2 (ab92494) at 0.5µg/ml.