Anti-SOX2 抗体

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX2 antibody (AB97959)

IHC image of SOX2 staining in Human brain glioma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab97959, 1μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

PubMed

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody (AB97959)

ICC/IF analysis of dissociated induced pluripotent stem cells from mouse embryonic fibroblasts stained for SOX-2 (Red) using ab97959. TOPRO (purple). Scale bar=100 μm

Muratore CR et al. Comparison and Optimization of hiPSC Forebrain Cortical Differentiation Protocols. PLoS One 9:e105807 (2014). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody (AB97959)

Cell line : NCCIT (human pluripotent embryonal carcinoma)

Target AbID : ab97959 anti-Sox2, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody was used.

Counterstain AbID : ab7291 anti-Tubulin (Rabbit mAb), 97959

Fixative : 4% PFA

Permeabilisation : 0.1% Triton-X

Nuclear counter stain : DAPI

Comments : Confocal image showing negative staining on NCCIT cells

Target primary antibody dilution : 1 : 500

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 : 500 (ab97959)

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)

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody (AB97959)

Cell line : NIH/3T3 (mouse embryonic fibroblast cell line)

Target AbID : ab97959 anti-Sox2, used Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody

Counterstain AbID : ab7291 anti-Tubulin (Rabbit 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 NIH/3T3 cells

Target primary antibody dilution : 1 : 500

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 : 500 (ab97959)

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)

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody (AB97959)

ab97959 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 ab97959 at 0.1μg/ml 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.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-SOX2 antibody (AB97959)

ab97959 staining SOX2 in mES cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween 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 ab97959 at 1µg/ml 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).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Ap22478****

Western blot - Anti-SOX2 antibody (AB97959)

Green signal from target - ab97959 observed at 40 kDa
Red signal from loading control ab9484 (GAPDH) observed at 37 kDa

All lanes:

Western blot - Anti-SOX2 antibody (ab97959) at 1 µg

Lane 1:

NCCIT (Human embryonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

F9 (Mouse embryonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

C6 (Rat glial tumor cell line) Whole Cell Lysate at 10 µg

Lane 5:

Hippocampus (Rat) Tissue Lysate at 10 µg

Lane 6:

Spinal Cord (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Blots were developed with goat anti-rabbit IgG (H+L) and goat anti-mouse IgG (H+L) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

Predicted band size: 34 kDa

Observed band size: 40 kDa

false

• WB

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Western blot - Anti-SOX2 antibody (AB97959)

All lanes:

Western blot - Anti-SOX2 antibody (ab97959) at 1 µg/mL

Lane 1:

IOUD2 (Mouse embryonic stem cell) Whole Cell Lysate at 10 µg

Lane 2:

HUES7 (Human embryonic stem cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 34 kDa

Observed band size: 37 kDa,39 kDa,43 kDa

true

Exposure time: 3min

• WB

Unknown

Western blot - Anti-SOX2 antibody (AB97959)

Secondary antibody - goat anti-rabbit HRP preadsorbed (ab97080)

All lanes:

Western blot - Anti-SOX2 antibody (ab97959) at 1/1000 dilution

All lanes:

Western blot - Recombinant Human SOX2 protein (<a href='/products/proteins-peptides/recombinant-human-sox2-protein-ab79950'>ab79950</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 34 kDa

true

Exposure time: 30s