Recombinant Anti-SOX2 antibody [EPR3131] - BSA and Azide free (ab215970)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab92494)

ab92494 staining SOX2 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% paraformaldehyde (10 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 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.

Imaging Mass Cytometry™ (IMC™) image of human glioblastoma brain cancer tissue stained with Anti-SOX2 antibody [EPR3131]. ab215970 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.

Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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.<\p>

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92494).

This ICC data was generated using the same anti-SOX2 antibody clone, EPR3131, in a different buffer formulation (cat# ab92494).

Cell line: F9 (mouse embryonal carcinoma)

Target AbID: Ab92494 anti-Sox2, Ab150077 AlexaFluor®488 Goat anti-Rabbit secondary

Counterstain AbID: Ab7291 anti-Tubulin (Mouse mAb), Ab150120 AlexaFluor®594 Goat anti-Mouse secondary

Fixative: 4% PFA

Permeabilisation: 0.1% Triton-X

Nuclear counter stain: DAPI

Comments: Confocal image showing negative staining on F9 cells

Target primary antibody dilution: 1:200

Target secondary antibody dilution: 1:1000 (2ug/mL)

Counterstain primary antibody dilution: 1:1000 (1ug/mL)

Counterstain secondary antibody dilution: 1:1000 (2ug/mL)

Negative control 1 primary antibody dilution: 1:200 (1ug/mL)  (Ab92494)

Negative control 1 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150120)

Negative control 2 primary antibody dilution: 1:1000 (1ug/mL) (Ab7291)

Negative control 2 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150077)