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Western blot
Flow cytometryAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- MA5-16399 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SOX2 Monoclonal Antibody (SP76)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is prostate or lung squamous cell carcinoma.
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- SP76
- Vial size
- 500 µL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references CDX2-induced intestinal metaplasia in human gastric organoids derived from induced pluripotent stem cells.
Autologous treatment for ALS with implication for broad neuroprotection.
Koide T, Koyanagi-Aoi M, Uehara K, Kakeji Y, Aoi T
iScience 2022 May 20;25(5):104314
iScience 2022 May 20;25(5):104314
Autologous treatment for ALS with implication for broad neuroprotection.
Kim D, Kim S, Sung A, Patel N, Wong N, Conboy MJ, Conboy IM
Translational neurodegeneration 2022 Mar 11;11(1):16
Translational neurodegeneration 2022 Mar 11;11(1):16
No comments: Submit comment
Supportive validation
- 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 (SP76) (Product # MA5-16399, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b and Fig. c). 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 (SP76) (Product # MA5-16399). A 38 kDa and 27 kDa band corresponding to SOX2 was observed across NTERA-2 and NCCIT. Nuclear enriched extracts (30 µg lysate) of NTERA-2 cl.D1 (Lane 1), HeLa (Lane 2), THP-1 (Lane 3) and NCCIT (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of MCF7 Cells using anti-SOX-2 Monoclonal Antibody (Product # MA5-16399). The recommened dilution for this antibody in western blot applications is 1:25.
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 (SP76) (Product # MA5-16399) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 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 localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SOX-2 using anti-SOX-2 Monoclonal Antibody (Product # MA5-16399) in New Born Brain Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) assay of endogenous SOX2 protein using Anti-SOX2 Antibody: ChIP was performed using Anti-SOX2 Monoclonal Antibody (Product # MA5-16399, 5 µl) on sheared chromatin from NTERA-2 cells using the MAGnify ChIP System (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over NANOG promoter (active), OCT4 promoter (active), SAT2 satellite repeats and GAPDH TSS (inactive). Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- The expression of the master transcription factors CDX1, CDX2, and SOX2 (A) RPKM values (log2) of CDX1, CDX2, and SOX2 in RNA-seq data. (B) Immunofluorescence analyses of E-Cadherin, SOX2, and CDX2 in gastric organoids from CDX2-iPSC at Day 40 without (-) (left panel) or with (+) (right panel) DOX treatment for 5 days, using confocal microscopy. Scale bars, 20 mum.
