Anti-FOXA2 antibody [EPR4465] - ChIP Grade

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

Immunofluorescence staining of HepG2 cells with purified ab108396 at a working dilution of 1 in 300, counter-stained with DAPI. Tubulin was stained with mouse anti-tubulin at a dilution of 1/1000 (ab7291) and Alexa Fluor^® 594 goat anti-mouse at a dilution of 1/500 (ab150120) . The secondary antibody was ab150077 Alexa Fluor^® 488 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in the bottom middle and right hand panels - for the first negative control, purified ab108396 was used at a dilution of 1/200 followed by an Alexa Fluor^® 555 goat anti-mouse antibody at a dilution of 1/500 and for the second negative control mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab15007) were used.

• ChIP

Unknown

ChIP - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

Chromatin was prepared from HepG2 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab108396 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

• WB

Unknown

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (ab108396) at 1/1000 dilution

All lanes:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 110 kDa,48 kDa

Observed band size: 52 kDa,98 kDa

false

• WB

Unknown

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (ab108396) at 1/5000 dilution

All lanes:

SW480 cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 22 kDa,48 kDa,72 kDa

Observed band size: 52 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab108396 [EPR4465]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab108396 [EPR4465]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab108396 [EPR4465]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Unknown

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (AB108396)

All lanes:

Western blot - Anti-FOXA2 antibody [EPR4465] - ChIP Grade (ab108396) at 1/1000 dilution

Lane 1:

HepG2 lysate at 10 µg

Lane 2:

Human colon cancer lysate at 10 µg

Predicted band size: 47 kDa,48 kDa

false