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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
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- Product number
- MA5-15449 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SOX2 Monoclonal Antibody (10F10C9)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15449 targets SOX2 in WB applications and shows reactivity with Human samples.
- Antibody clone number
- 10F10C9
- Concentration
- Conc. Not Determined
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SOX2 using a SOX2 monoclonal antibody (Product # MA5-15449) against a truncated Trx-SOX2 recombinant protein (1) and truncated MBP-SOX2 (aa1-170) recombinant protein (2).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SOX2 was achieved by transfecting NTERA-2 cl.D1 with SOX2 specific siRNAs (Silencer® select Product # S13294, S13296). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the SOX2 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SOX2 Monoclonal Antibody (10F10C9) (Product # MA5-15449, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:2000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SOX2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SOX2 Monoclonal Antibody (10F10C9) (Product # MA5-15449) and a 27 kDa band corresponding to SOX2 was observed in HEK-293 and NTERA-2. Nuclear enriched extracts (30 µg lysate) of HEK-293 (Lane 1), NTERA-2 cl.D1 (Lane 2), HeLa (Lane 3) and THP-1 (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SOX2 was performed using 70% confluent log phase NTERA-2 cl.D1 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SOX2 Monoclonal Antibody (10F10C9) (Product # MA5-15449) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766, 1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
