Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal

• ICC

Supplier Data

Immunocytochemistry - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT1080 (Human fibrosarcoma cell line) cells labeled for SATB2 using ab51502 at 100 μg/ml in immunocytochemistry.

• IP

Supplier Data

Immunoprecipitation - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

Immunoprecipitation using ab51502 at 500 μg/sample.

Sample : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate prepared in RIPA buffer.

All lanes:

Immunoprecipitation - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (ab51502)

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• WB

Lab

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

False colour image of Western blot : Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal staining at 2 µg/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20,000 dilution, shown in red.

In Western blot, ab51502 was shown to bind to recombinant SATB1 and recombinant SATB2 protein. A band was observed at 100 kDa in wild-type HAP1 cell lysates with no signal observed at this size in SATB2 knockout cell line. To generate this image, wild-type and SATB2 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (ab51502) at 2 µg

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

SATB2 knockout HAP1 cell lysate at 20 µg

Lane 3:

Mouse E18 embyonic brain tissue lysate at 20 µg

Lane 4:

NIH/3T3 cell lysate at 20 µg

Lane 5:

HT1080 cell lysate at 20 µg

Lane 6:

Saos-2 cell lysate at 20 µg

Lanes 7 and 9:

Empty

Lane 8:

SATB1 recombinant protein at 0.1 µg

Lane 10:

SATB2 recombinant protein (GST-tag) (<a href='/en-us/products/proteins-peptides/recombinant-human-satb2-protein-ab132405'>ab132405</a>) at 0.1 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 100 kDa

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• WB

Lab

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

False colour image of Western blot : Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal staining at 2 µg/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20,000 dilution, shown in red.

In Western blot, ab51502 was shown to bind to recombinant SATB1 and recombinant SATB2 protein. A band was observed at 100 kDa in wild-type HAP1 cell lysates with no signal observed at this size in SATB2 knockout cell line. To generate this image, wild-type and SATB2 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (ab51502) at 2 µg

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

SATB2 knockout HAP1 cell lysate at 20 µg

Lane 3:

Mouse E18 Embyonic brain cell lysate at 20 µg

Lane 4:

NIH 3T3 cell lysate at 20 µg

Lane 5:

HT1080 cell lysate at 20 µg

Lane 6:

Saos-2 cell lysate at 20 µg

Secondary

All lanes:

Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

All lanes:

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (ab51502) at 2 µg/mL

Lane 1:

HT1080 (Human fibrosarcoma cell line) whole cell lysate at 25 µg

Lane 2:

NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 25 µg

Observed band size: 82 kDa

false

• WB

CiteAb

Western blot - Anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal (AB51502)

SATB1 + SATB2 western blot using anti-SATB1 + SATB2 antibody [SATBA4B10] - C-terminal ab51502. Publication image and figure legend from Wong, F. K., Fei, J. F., et al., 2015, PLoS Biol, PubMed 26252244.

ab51502 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab51502 please see the product overview.

Conditional Pax6 expression in Tis21-positive cortical progenitors increases upper-layer neuron production.(A) Flow scheme of the experiment. (B–J) Dorsolateral telencephalon of tamoxifen-treated E14.5 (D,E) and E17.5 (B–J) Tis21–CreERT2 heterozygous mice electroporated at E13.5 with control (B,D–F,G,I,J) or Pax6-expressing (C–F,H–J) plasmid and subjected to a single EdU pulse at 10 h after electroporation (E14.0). (B,C,G,H) Tbr1 (B,C) and Satb2 (G,H) immunofluorescence (blue) and EdU (white) and RFP (red) fluorescence in the cortical plate (coronal 50-μm vibratome sections). Insets are representative examples of RFP-positive nuclei (outlined by dashed yellow lines) at higher magnification that are either Tbr1- or Satb2-positive. Scale bars, 20 μm (B,C,G,H) and 5 μm (insets in B,C,G,H). (D,E) Quantification of RFP+ nuclei in the entire cortical wall (200-μm wide area) (D) and their fold-increase at E17.5 as compared to E14.5 (E), upon control (Con, white) and Pax6 (black) electroporation. (D) E14.5, mean of three independent experiments, each being the average of two or three embryos (eight in total); E17.5, mean of eight embryos from at least two independent experiments; (E) mean of eight E17.5/E14.5 ratios. (F,I,J) Quantification of Tbr1+, EdU+ and RFP+ triple-positive nuclei (F), Satb2+, EdU+ and RFP+ triple-positive nuclei (I) and Brn2+ and RFP+ double-positive nuclei (J) in the cortical plate, expressed as percentage of all cells that are EdU+ and RFP+ (F,I) and RFP+ (J) in the cortical plate (200-μm wide area) upon control (Con, white) and Pax6 (black) electroporation. (F,I) Mean of three independent experiments, each being the average of two to three embryos. (J) Mean of four embryos from two different litters. Error bars, SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

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