MA1-014

antibody from Invitrogen Antibodies

Targeting: SOX2

Provider product page for MA1-014

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Chromatin Immunoprecipitation

Other assay

Antibody data

Antibody Data
Antigen structure
References [22]
Comments [0]
Validations
Immunocytochemistry [3]
Flow cytometry [3]
Chromatin Immunoprecipitation [1]
Other assay [10]

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Product number: MA1-014 - Provider product page
Provider: Invitrogen Antibodies
Product name: SOX2 Monoclonal Antibody (20G5)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: Western blot analysis of MA1-014 specifically detects SOX2 protein at ~36 kDa in the nucleus of human and mouse embryonal carcinoma and iPS cells.
Reactivity: Human, Mouse, Canine
Host: Mouse
Isotype: IgG
Antibody clone number: 20G5
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20°C

SOX2 protein structure - MA1-014 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Sox2 (green) in HEL 11.4 induced IPS cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a FITC-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Sox2 (green) in H9 embryonic stem cells grown for a few days on Matrigel-coated chamber slides. Cells fixed in 4% paraformaldehyde were permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were probed with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:200 overnight at 4°C, washed with PBST, and incubated with a FITC-conjugated secondary antibody at a dilution of 1:100 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI and cells were analyzed by fluorescence microscopy at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sox2 (green) in human neural stem cells derived from PD-3 iPSCs using Gibco® PSC Neural Induction Medium (Product # A1647801). The cells were fixed and permeabilized using Image-IT® Fixation/Permeabilization kit (Product # R37602), and blocked with blocking buffer included the kit for one hour at room temperature. Cells were stained with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:200 in blocking buffer for 3 hours at room temperature, and then incubated with an DyLight 488- conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:250 for 1 hour at room temperature. Image was taken on an EVOS® FLoid® Cell Imaging Station at 10X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Sox2 on human neural stem cells derived from PD-3 iPSCs using Gibco® PSC Neural Induction Medium (Product # A1647801). Cells were fixed, permeabilized and stained with a Sox2 monoclonal antibody (Product # MA1-014, red histogram) or mouse IgG1 isotype control (black histogram) at a 1:100 dilution in 5% BSA. After incubation of the primary antibody for 1 hour on ice, the cells were stained with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:500 for 1 hour on ice. A representative 10,000 cells were acquired for each sample.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometric analysis of Sox2 (blue histogram) on HEL 11.4 induced IPS cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37°C. Cells were incubated with a Sox2 monoclonal antibody (Product # MA1-014) or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a FITC-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500 µL of FACS buffer containing 10 µL of 4% paraformaldehyde, and analyzed on a flow cytometer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometric analysis of Sox2 (blue histogram) on H9 embryonic stem cells. To generate single cells suspensions, colonies were treated with TrypLE cell dissociation enzyme for 5 minutes at 37°C. Cells were incubated with a Sox2 monoclonal antibody (Product # MA1-014) or mouse IgG (green histogram) at a dilution of 1:100 for 1 hour on ice, washed with PBS + 5% fetal calf serum (FACS buffer), and incubated with a FITC-conjugated secondary antibody at a dilution of 1:200 for 30 minutes on ice. Cells were washed with cold FACS buffer, resuspended in 500 µL of FACS buffer containing 10 µL of 4% paraformaldehyde, and analyzed on a flow cytometer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of Sox2 was performed on NCCIT cells. Antigen-antibody complexes were formed by incubating 500 µg of NCCIT whole cell lysate with 5 µg of a Sox2 monoclonal antibody (Product # MA1-014) overnight on a rocking platform at 4øC. The immune complexes were captured on 50 µL Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Eluted sample and 25 µg of NCCIT whole cell lysate (loading control) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween-20 for at least 1 hour. The membrane was probed with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:1000 overnight rotating at 4øC, washed in TBST, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 32430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3. Analysis of pluripotency transcription factors in response to ETO treatment in PA-1 cells. ( A ) Protein expression of p53, p21Cip1, OCT4A, SOX2 and NANOG were analyzed by immunoblotting at indicated time points after ETO-treatment. GAPDH was used as a loading control NT2 cells were used as a positive control for pluripotency genes OCT4A, SOX2 and NANOG. p53 and p2Cip1 protein expression was upregulated in the DNA damage response. An upregulation of the OCT4A protein was detected from day 1 post ETO treatment, while SOX2 remained low and unchanged post ETO expression. NANOG was not detected in NT or ETO treated PA-1 cells. Results are representative of 3 independent experiments. ( B ) Semi-automatic image cytometry of SOX2 expression was performed on the nuclei of individual cells. Box-plots indicate the heterogeneity and expression levels measured. The data show that SOX2 remains unresponsive to ETO-treatment.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Immunohistochemical analysis of SOX-2 positive cells in five cases of pneumocytoma of the lung. Tissue sections were stained with anti-SOX-2 antibody to detect the positivity in these five different cases of pneumocytoma of the lung. (A-O) Case 1, (B-P) Case 2, (C-Q) Case 3, (D-R) Case 4, (E-S) Case 5. Semi-quantitative method was used to assess the SOX-2 positivity of the tumor cells: 0 (25-50% positive), 3 (>50-75% positive), and 4 (>75% positive). Representative images of the five cases are shown at 10, 20, and 40x magnification. Isotype control for SOX-2 antibody was shown (F, N, T) . Scale bar = 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 Gene expression and protein immunofluorescence analysis of 3D sarcospheres scaffold-based models. Relative quantification of OCT-4, NANOG and SOX-2, stemness marker genes, SAOS-2 ( a ) and MG63 ( b ). Relative quantification of HIF-1a, NOTCH-1 and IL-6, CSCs niche-related genes, SAOS-2 ( c ) and MG63 ( d ). The graphs show the fold change expression of the genes relative to the scaffold-free sarcospheres at day 10 of culture (mean +- standard error; * p value

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Evaluation of cellular identity and morphology in 3D brain organoid. A Representative immunostaining showing the presence of a N-CADHERIN-positive (green) germinal layer in correspondence of the apical membrane of the ventricular zone layer in a FMRP-WT brain organoid at day 50. B , C Representative images revealing forebrain progenitor markers in developing cortical plates within FMRP-WT brain organoid at day 50: B SOX2 (green), DAPI (blue); C PAX6 (red), FOXG1 (green) and DAPI (blue). D Representative staining of early-born deep-layer cortical neurons positive for TBR1 (red), CTIP2 (green) markers, and pan-neuronal MAP2 marker (white) in a FMRP-WT brain organoid at day 50. E Representative images of a cortical plate from a FMRP-WT brain organoid at day 100 revealing heterogeneous cellular population, labeled for specific cortical neuronal marker (CTIP2, green), astrocytic marker (GFAP, red), and neuronal progenitor marker (PAX6, white). Nuclei were stained with DAPI (blue). Scale bar: 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 SOX2-SOX9 status for a range of different seeding densities. Fluorescent micrographs of day 17 lung progenitors seeded for 8 days in 100 um tubes and stained for SOX9 (red) and SOX2 (white) are single positive for SOX9 in 200,000 cells/cm 2 ( A ), 300,000 cells/cm 2 ( B ), 500,000 cells/cm 2 ( C ), 600,000 cells/cm 2 ( D ), 800,000 cells/cm 2 ( E ), 1,000,000 cells/cm 2 ( F ). Bar charts showing quantification of the percentage of SOX9 + cells and the percentage of SOX9 + SOX2 + cells in 100 um tubes ( G ) and 1400 um tubes ( H ) for a range of seeding densities. A significant increase in SOX9 single positive cells and a significant decrease in SOX2 + SOX9 + was observed at all densities in 100-micron diameter tubes; * p < 0.05. Bar, 50 um; the dotted white line approximates the PDMS border; n = 3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Expression of stemness markers: immunocytochemistry images of ( A ) Sox2 primary antibody stained with FITC and counterstained with DAPI; ( B ) Nestin-primary antibody stained with PE and counterstained with DAPI. The scale bar indicates 20 um.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Immunohistochemistry of lung myofibroblastic tumor. Representative immunohistochemical staining of ALDH1A1, SOX2, NANOG, OCT-4, and c-MYC stem cell markers on a myofibroblastic tumor. Images were shown at 10x and 20x magnification.