Anti-ATF2 (phospho T71) antibody [E268] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ATF2 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

Immunocytochemistry of HeLa (Human epithelial cell line from cervix adenocarcinoma), prepared in FBS free medium overnight labeling ATF2 at 0.9 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Alexa Fluor^® 488 Goat anti-Rabbit (ab150077) was used as the secondary antibody at 1/500. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used as the counter stain at 2.5 μg/ml. DAPI was used for nuclear counter stain. Confocal image showing the expression was increased on HeLa cells, prepared in FBS free medium overnight, then treated with 250ng/ml anisomycin for 30min.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32019).

• IP

Supplier Data

Immunoprecipitation - Anti-ATF2 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

ab32019 Immunoprecipitating ATF2 in Human Hela whole cell lysate. For western blotting ab32019 (1 : 1000) was used to confirm successful immunoprecipitation. Blocking and diluting buffer used was 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32019).

All lanes:

Immunoprecipitation - Anti-ATF2 (phospho T71) antibody [E268] (<a href='/en-us/products/primary-antibodies/atf2-phospho-t71-antibody-e268-ab32019'>ab32019</a>) at 0.47 µg/mL

Lane 1:

HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate (input) at 10 µg

Lane 2:

HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate (+)

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/atf2-phospho-t71-antibody-e268-ab32019'>ab32019</a> in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate (-)

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/10000 dilution

Predicted band size: 55 kDa

false

Exposure time: 3min

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-ATF2 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

This data was developed using the same antibody clone in a different buffer formulation (ab32019).

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 treated with anisomycin (25 µg/mL 30 min) and 5 µg of ab ab32019[E268]. 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 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

This data was developed using the same antibody clone in a different buffer formulation (ab32019).

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 treated with anisomycin (25 µg/mL 30 min) and 5 µg of ab ab32019[E268]. 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 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

This data was developed using the same antibody clone in a different buffer formulation (ab32019).

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 treated with anisomycin (25 µg/mL 30 min) and 5 µg of ab ab32019[E268]. 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.

• Dot

Lab

Dot Blot - Anti-ATF2 (phospho T71) antibody [E268] - BSA and Azide free (AB242381)

Blocking and diluting buffer used was 5% NFDM/TBST.

Primary : ab32019, 0.5 ug/ml
Secondary : ab97051, 1/100000 dilution