Anti-GATA3 antibody [L50-823]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GATA3 antibody [L50-823] (AB282110)

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling GATA3 with ab282110 at 1/2000 (0.533 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining in human breast cancer. The section was incubated with ab282110 for 30 minutes 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GATA3 antibody [L50-823] (AB282110)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling GATA3 with ab282110 at 1/50 (21.32 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in MCF7 cells.

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GATA3 antibody [L50-823] (AB282110)

ab282110 staining GATA3 in MCF7 cells, with negative expression in HepG2 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab282110 at 10 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin at 1 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GATA3 antibody [L50-823] (AB282110)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GATA3 with ab282110 at 1/2000 (0.533 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining in rat spleen. The section was incubated with ab282110 for 30 minutes 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GATA3 antibody [L50-823] (AB282110)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GATA3 with ab282110 at 1/2000 (0.533 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Nuclear staining in mouse spleen. The section was incubated with ab282110 for 30 minutes 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 minutes.

• WB

Lab

Western blot - Anti-GATA3 antibody [L50-823] (AB282110)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24758297).

Exposure time : Lane 1 : 70 seconds

Lane 2 : 3 seconds.

All lanes:

Western blot - Anti-GATA3 antibody [L50-823] (ab282110) at 1/1000 dilution

Lane 1:

T47D (human ductal breast epithelial tumor epithelial cell) nuclear lysate at 10 µg

Lane 2:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GATA3 antibody [L50-823] (AB282110)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24758297).

Low expression : MDA-MB-231(PMID : 31685800)

Exposure time : 10 seconds.

All lanes:

Western blot - Anti-GATA3 antibody [L50-823] (ab282110) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 2:

HT-1376 (Human urinary bladder carcinoma cell line) whole cell lysate at 10 µg

Lane 3:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 10 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 5:

MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 47 kDa

Observed band size: 37 kDa,50 kDa

false