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Recombinant full length protein corresponding to Human SOX2.
Our Abpromise guarantee covers the use of ab75485 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC - Wholemount||Use at an assay dependent concentration.|
|Flow Cyt||1/10 - 1/50. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|Sandwich ELISA||Use a concentration of 0.2 µg/ml. Can be paired for Sandwich ELISA with Rabbit monoclonal [EPR3131] to SOX2 (ab92494).
For sandwich ELISA, use this antibody as Capture at 0.2 µg/ml with Rabbit monoclonal [EPR3131] to SOX2 (ab92494) as Detection.
|WB||1/200 - 1/2000. Predicted molecular weight: 34 kDa.|
|IHC-P||1/50 - 1/100.|
Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemical staining of SOX2 in Human lung carcinoma tissue sections (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections) with ab75485 at a dilution of 1/25. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C. Antigen retrieval was heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A HRP-conjugated goat anti-mouse polyclonal (ready to use) was used as the secondary antibody.
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 25°C. 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.
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 at 24°C. Primary antibody used at 1 µg/ml incubated for 24 hours at 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.
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 +4°C. 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.
Immunocytochemistry/Immunofluorescence analysis of A549 cells labelling SOX2 with ab75485 at 1/25
A549 cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.1%, 10 min), then incubated with the primary antibody (1/25, 1 h at 37°C). Alexa Fluor® 488 conjugated donkey anti-mouse antibody (green) was used (1/400, 50 min at 37°C). Cytoplasmic actin was counterstained with Alexa Fluor® 555 (red) conjugated Phalloidin (1 h at 37°C). SOX2 immunoreactivity is localized to nucleus significantly.
Immunocytochemistry/Immunfluorescence analysis of Sy5Y cells labelling SOX2 with ab75485 at 1/100.
SY5Y cells were fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with the primary antibody (1/100, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-mouse antibody (green) was used (1/1000, 1h). Note the highly specific localization of the SOX2 mainly in the nucleus.
Overlay histogram showing F9 cells stained with ab75485 (red line). The cells were fixed with 80% methanol (5 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 (ab75485, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse 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 4% paraformaldehyde (10 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"