Anti-ATF2 antibody [E243] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using ab32160, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATF2 (red) with ab32160 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using ab32160, the same antibody clone in a different buffer formulation.

ab32160, at a dilution of 1/250, staining ATF2 in paraffin embedded breast carcinoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using the same antibody clone in a different buffer formulation (ab32160).
Immunocytochemistry analysis of A549 (human lung carcinoma epithelial cell) labeling ATF2 with purified ab32160 at 1/100 dilution (10 µg/ml). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.10 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control : PBS instead of the primary antibody.

• ChIP

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ChIP - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using ab32160, the same antibody clone in a different buffer formulation.

Chromatin was prepared from Jurkat (TPA and Ionomycin treated or not) cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 5μg of ab32160 (red), and 20μl of protein A/G sepharose beads slurry (10μl of sepharose A beads + 10μl of sepharose G beads). 5μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

• IP

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Immunoprecipitation - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using ab32160, the same antibody clone in a different buffer formulation.

ab32160 (purified) at 1/60 dilution (20 μg/mL) immunoprecipitating ATF2 in HeLa whole cell lysate.
Lane 3 (-) : HeLa(Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μgab32160 & HeLa whole cell lysateRabbit monoclonal IgG (ab172730) instead of ab32160 in HeLa whole cell lysate
For western blotting, ab32160 at 1/1000 dilution (2.284 μg/mL) and
bbit TureBlot : Anti-Rabbit IgG HRP was used as the secondary antibody at 1/1500 dilution.
Blocking and diluting buffer : 5% NFDM /TBST .

All lanes:

Immunoprecipitation - Anti-ATF2 antibody [E243] - ChIP Grade (<a href='/en-us/products/primary-antibodies/atf2-antibody-e243-chip-grade-ab32160'>ab32160</a>)

Predicted band size: 55 kDa

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• WB

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Western blot - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using ab32160, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ATF2 antibody [E243] - ChIP Grade (<a href='/en-us/products/primary-antibodies/atf2-antibody-e243-chip-grade-ab32160'>ab32160</a>) at 1/10000 dilution

All lanes:

HeLa cell lysate

Predicted band size: 55 kDa

Observed band size: 70 kDa

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• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using the same antibody clone in a different buffer formulation (ab32160).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab ab32160[E243]. 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-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using the same antibody clone in a different buffer formulation (ab32160).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab ab32160[E243]. 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-ATF2 antibody [E243] - BSA and Azide free (AB247240)

This data was developed using the same antibody clone in a different buffer formulation (ab32160).

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ K-562 (human chronic myelogenous leukemia lymphoblast) cells and 5 µg of ab ab32160[E243]. 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.