Sandwich ELISA - SOX2 antibody [57CT23.3.4] (ab75485)

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.

Western blot - SOX2 antibody [57CT23.3.4] (ab75485)

Anti-SOX2 antibody [57CT23.3.4] (ab75485) at 1/200 dilution + SOX2 recombinant protein

Predicted band size : 34 kDa
Observed band size : 38 kDa (why is the actual band size different from the predicted?)

Western blot - SOX2 antibody [57CT23.3.4] (ab75485)

All lanes : Anti-SOX2 antibody [57CT23.3.4] (ab75485) at 1/200 dilution

Lane 1 : non-transfected 293 cell lysate
Lane 2 : lysate of 293 cells transiently transfected with SOX2 gene

Lysates/proteins at 12.5 µg per lane.


Predicted band size : 34 kDa
Observed band size : 36 kDa (why is the actual band size different from the predicted?)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - SOX2 antibody [57CT23.3.4] (ab75485)

Human lung carcinoma tissue with ab75485 at 1/50 dilution, using a peroxidase-conjugated secondary antibody, followed by DAB staining.

Immunocytochemistry/ Immunofluorescence - SOX2 antibody [57CT23.3.4] (ab75485)

ab75485 staining SOX2 in human schwann cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in 4% paraformaldehyde for 10 minutes, permeabilised in 0.2% Triton X-100 and then blocked using 10% DCS/ 0.1M lysine/PBS for 30 minutes at 25ºC. Samples were then incubated with primary antibody at 1/100 for 16 hours at 4°C. The secondary antibody used was a rabbit anti-mouse IgG conjugated to Alexa Fluor® 568 used at a 1/500 dilution. Cells were counterstained with Hoechst dye to show nuclei.

This image was kindly supplied by Dr David Parkinson by Abreview

Immunocytochemistry/ Immunofluorescence - SOX2 antibody [57CT23.3.4] (ab75485)

Immunocytochemistical detection of SOX2 antibody (ab75485) on a mixed rat glial cell culture. Fixative: Paraformaldehyde Permeabilization: 0.3x Triton. Blocking step: 10% serum for 1 hour @ 24°C. Primary antibody used at 1 µg/ml incubated for 24 hours @ 4°C. Secondary antibody: Alexa Fluor® 568 (1/1000). The submitted image shows rat primary glial culture stained with ab75485; DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom

Immunocytochemistry/ Immunofluorescence - SOX2 antibody [57CT23.3.4] (ab75485)

ICC/IF image of ab75485 stained mouse embryonic stem cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75485, 5µg/ml) overnight at +4ºC. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.