Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-OV-3(human ovarian cancer epithelial cell) cells labelling Gata6 with ab322895 at 1/100 (5.25 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in SK-OV-3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : HL-60.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human ovarian cancer tissue labeling Gata6 with ab322895 at 1/500 (1.05 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human ovarian cancer.
The section was incubated with ab322895 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human squamous cell lung carcinoma  tissue labeling Gata6 with ab322895 at 1/500 (1.05 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human squamous cell lung carcinoma.
The section was incubated with ab322895 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Gata6 with ab322895 at 1/500 (1.05 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.
The section was incubated with ab322895 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Gata6 with ab322895 at 1/500 (1.05 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung.
The section was incubated with ab322895 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling Gata6 with ab322895 at 1/500 (1.05 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human gastric adenocarcinoma.
The section was incubated with ab322895 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0 Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Caco-2 (human colorectal adenocarcinoma epithelial cell) cells labelling Gata6 with ab322895 at 1/1000 (0.525 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in Caco-2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Huh7 (human hepatocellular carcinoma epithelial cell) cells labelling Gata6 with ab322895 at 1/100 (5.25 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).

Confocal image showing nuclear staining in Huh7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : THP-1.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Chromatin was prepared from Caco-2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab322895 [EPR29847-53]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Chromatin was prepared from Caco-2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab322895 [EPR29847-53]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Chromatin was prepared from Caco-2 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 ug of ab322895 [EPR29847-53]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Supplier Data

Immunoprecipitation - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Gata6 was immunoprecipitated from 0.35 mg Caco-2 whole cell lysate with ab322895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322895 at 1/1000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Gata6 antibody [EPR29847-53] (<a href='/en-us/products/primary-antibodies/gata6-antibody-epr29847-53-ab322895'>ab322895</a>) at 1/1000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/gata6-antibody-epr29847-53-ab322895'>ab322895</a> IP in Caco-2 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/gata6-antibody-epr29847-53-ab322895'>ab322895</a> in Caco-2 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 1/5000 dilution

Observed band size: 60 kDa,75 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HL-60

The identity of the bands lower than 60 kDa are unknown.

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Gata6 antibody [EPR29847-53] (<a href='/en-us/products/primary-antibodies/gata6-antibody-epr29847-53-ab322895'>ab322895</a>) at 1/1000 dilution

Lane 1:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 2:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 3:

Human colon tissue lysate at 20 µg

Lane 4:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 60 kDa,75 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : THP-1

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 27273097).

In Western blot Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Gata6 antibody [EPR29847-53] (<a href='/en-us/products/primary-antibodies/gata6-antibody-epr29847-53-ab322895'>ab322895</a>) at 1/1000 dilution

Lane 1:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa,75 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands lower than 60 kDa are unknown.

All lanes:

Western blot - Anti-Gata6 antibody [EPR29847-53] (<a href='/en-us/products/primary-antibodies/gata6-antibody-epr29847-53-ab322895'>ab322895</a>) at 1/1000 dilution

All lanes:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60 kDa,75 kDa

false

Exposure time: 180s

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 ug of ab322895 [EPR29847-53]. 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

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 ug of ab322895 [EPR29847-53]. 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

Supplier Data

ChIC/CUT&RUN sequencing - Anti-Gata6 antibody [EPR29847-53] - BSA and Azide free (AB326609)

This data was developed using ab322895, the same antibody clone in a different buffer formulation.

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL 2.5 x 10⁵ Caco-2 (human colorectal adenocarcinoma epithelial cell) cells and 5 ug of ab322895 [EPR29847-53]. 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.