Anti-CEBP Alpha/CEBPA antibody [EP708Y]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

Overlay histogram showing HeLa cells stained with ab40761 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40761, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

ab40761 (1/250) staining HeLa cells using immunofluorescence.

• WB

AbReview29648****

Western blot - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

Blocked with 5% Milk for 1 hour at 24°C.

We are unsure as to the identity of the 30kda band.

All lanes:

Western blot - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (ab40761) at 1/5000 dilution

All lanes:

Human MCF-7 whole cell lysate at 100000 Cells

Secondary

All lanes:

HRP-conjugated Donkey anti-rabbit IgG polyclonal at 1/2000 dilution

Predicted band size: 38 kDa

Observed band size: 40 kDa

true

Exposure time: 1min

This image is courtesy of an anonymous Abreview

• WB

Unknown

Western blot - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

We are unsure as to the identity of the 30kda band.

All lanes:

Western blot - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (ab40761) at 1/1000 dilution

All lanes:

U937 + LPS cell lysate at 10 µg

Predicted band size: 38 kDa

Observed band size: 30 kDa,43 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 THP-1 (human monocytic leukemia monocyte) cells and 5 µg of ab40761 [EP708Y]. 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-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 THP-1 (human monocytic leukemia monocyte) cells and 5 µg of ab40761 [EP708Y]. 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) method

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-CEBP Alpha/CEBPA antibody [EP708Y] (AB40761)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 THP-1 (human monocytic leukemia monocyte) cells and 5 µg of ab40761 [EP708Y]. 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.