- PA5-30162 - Invitrogen Antibodies SATB1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot analysis of SATB1 was performed by separating 50 µg of Various tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a SATB1 Polyclonal Antibody (Product # PA5-30162) at a dilution of 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SATB1 Polyclonal Antibody detects SATB1 protein by western blot analysis. Whole cell extracts (30 µg) was separated by 7.5 % SDS-PAGE, and blotted with SATB1 Polyclonal Antibody (Product # PA5-30162) diluted by 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of SATB1 was performed by separating 50 µg of Mouse tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a SATB1 Polyclonal Antibody (Product # PA5-30162) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: SATB1 Polyclonal Antibody detects SATB1 protein by western blot analysis. Whole cell extracts (30 µg) was separated by 7.5 % SDS-PAGE, and blotted with SATB1 Polyclonal Antibody (Product # PA5-30162) diluted by 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of SATB1 was achieved by transfecting IMR-32 cells with SATB1 specific siRNAs (Silencer® select Product # s12481, Product # s12479). Western blot analysis (Fig. a) was performed using membrane extracts from the SATB1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed using Laminin gamma-1 Polyclonal Antibody (Product # PA5-30162, 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). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SATB1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-SATB1 Polyclonal Antibody (Product # PA5-30162) and a ~100 kDa band corresponding to SATB1 was observed across the cell lines tested. Modified whole cell extracts (1% SDS) (30ug lysate) of IMR-32 (Lane 1), THP-1 (Lane 2), MCF-7 (Lane 3), MDA-MB-231 (Lane 4) and SK-N-SH (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of SATB1 was performed using 70% confluent log phase MDA-MB-231 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 10 minutes at room temperature. The cells were labeled with SATB1 Polyclonal Antibody (Product # PA5-30162) at 1:500 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with DAPI. F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded TOV-21G xenograft, using SATB1 (Product # PA5-30162) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: SATB1 Polyclonal Antibody detects SATB1 protein at nucleus on mouse fore brain by immunohistochemical analysis. Sample: Paraffin-embedded mouse fore brain. SATB1 Polyclonal Antibody (Product # PA5-30162) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: SATB1 Polyclonal Antibody detects SATB1 protein at nucleus on rat middle brain by immunohistochemical analysis. Sample: Paraffin-embedded rat middle brain. SATB1 Polyclonal Antibody (Product # PA5-30162) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Chromatin Immunoprecipitation (ChIP) assay of endogenous SATB1 protein using Anti-SATB1 Antibody: ChIP was performed using Anti-SATB1 Rabbit polyclonal Antibody (Product # PA5-30162, 5 µg) on sheared chromatin from IMR32 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to FOSB transcriptional start site, EGR2 intronic region (+2.5kb), ELAVL4 exonic region (+59kb) and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-30162