Anti-FOXA1 +FOXA2 抗体

• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

Lanes 1 - 4 : Merged signal (red and green). Green - ab23738 observed at 49 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab23738 was shown to recognize FOXA1 in wild-type HAP1 cells as signal was lost at the expected MW in FOXA1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FOXA1 knockout samples were subjected to SDS-PAGE. ab23738 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 0.7 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

HAP1 cells are predicted to express FOXA1, not FOXA2

All lanes:

Western blot - Anti-FOXA1 + FOXA2 antibody (ab23738) at 0.7 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

FOXA1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

A549 whole cell lysate at 20 µg

Predicted band size: 49 kDa

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• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

Western blot : Anti-FOXA1 antibody ab23738 staining at 0.7 µg/mL, shown in green; Mouse anti GST ab92 loading control staining at 1/1000 dilution, shown in magenta. A band was observed at 76 kDa. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FOXA1 + FOXA2 antibody (ab23738) at 0.7 µg/mL

Lane 1:

FOXA1 Recombinant Protein at 0.1 µg

Lane 2:

FOXA2 Recombinant Protein at 0.1 µg

Lane 3:

FOXA3 Recombinant Protein at 0.1 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 52 kDa

Observed band size: 76 kDa

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• WB

AbReview15258****

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

All lanes:

Western blot - Anti-FOXA1 + FOXA2 antibody (ab23738) at 1/1000 dilution

Lane 1:

Lysate prepared from human LNCaP cells at 20 µg

Lane 2:

Lysate prepared from MCF7 cells at 20 µg

Lane 3:

Lysate prepared from human LNCaP cells with ethanol as vehicle control at 20 µg

Lane 4:

Lysate prepared from human LNCaP cells with 100 nM DHT at 20 µg

Secondary

All lanes:

HRP-conjugated goat polyclonal to rabbit IgG at 1/5000 dilution

Predicted band size: 49 kDa

Observed band size: 51 kDa

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Exposure time: 2s

This image is a courtesy of Anonymous Abreview

• WB

Unknown

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

All lanes:

Western blot - Anti-FOXA1 + FOXA2 antibody (ab23738) at 1 µg/mL

All lanes:

Recombinant Human FOXA1 protein (<a href='/products/unavailable/recombinant-human-foxa1-protein-ab98301'>ab98301</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 49 kDa

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Exposure time: 10s

• WB

Unknown

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

All lanes:

Western blot - Anti-FOXA1 + FOXA2 antibody (ab23738) at 1 µg/mL

All lanes:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Predicted band size: 49 kDa

Observed band size: 31 kDa,51 kDa

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• WB

CiteAb

Western blot - Anti-FOXA1 + FOXA2 antibody (AB23738)

Western Blotting using Anti-FOXA1 antibody, ab23738. Publication image from Jiang, G. et al., 2019, Theranostics, 31588232. Legend direct from paper.

These factors are required for ESR1+ breast cancer. (A) Representation of known motif results for ER binding using the Homer software. (B) Expression profiles from primary breast tumours reveal that the mRNA levels of the three TFs are correlated with the ERα histological status and mRNA expression. The significant p-value is presented. (C) Western blotting experiments of corresponding TFs and hormone receptors were performed on a panel of nine breast cancer cell lines, including ER positive (MCF-7, T47-D, and ZR-75-1), AR positive (MDA-MB-453) and ERα-negative (SKBR3, SUM159, MDA-MB-435, MDA-MB-435, MDA-MB-231, and LM-2) cells. (D) The heatmap represents the ChIP-seq tag density for the FOXA1, GATA3, NR2F2, H3K4me1, and H3K27ac DNase-seq results obtained for the MCF-7 cells, ChIP-seq for H3K27ac and H3K4me1, and ATAC-seq for MDA-MB-231 cells at the defined groups characterized by the Venn diagram. (E) The average density distribution of H3K4me1 and H3K27ac among the seven different combinations of the three transcription factors in the MDA-MB-231 cell line. (F) Kaplan-Meier analysis of metastasis based on the GATA3, NR2F2, and FOXA1 expression levels in the patients. High transcript levels of the three transcription factors were correlated with improved lung metastasis-free survival (LMFS) in breast cancer patients. Analysis of eight groups established by the combination of the three expression levels showing that high expression of all three factors can synergistically improve LMFS. The TF order is FOXA1, NR2F2, and GATA3. For example, E110 indicates patients with high FOXA1 and NR2F2 expression relative to the average value and low GATA3 expression.

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