Immunofluorescent analysis of GATA1 (green) in U251 cells (right) using Rabbit anti GATA1 antibody compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with GATA1 polyclonal antibody in 3% BSA-PBS at a dilution of 1/100 and incubated overnight at 4 degree C in a humidified chamber. Cells were washed with PBST and incubated cells were counterstained with phalloidin (F-actin, red) and Hoechst 33342 (Nucleus, blue). Images were taken at a magnification of 60x

Immunohistochemistry analysis of GATA1 using Rabbit anti GATA1 antibody showing staining in the nucleus of paraffin-embedded human breast tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with GATA1 polyclonal antibody diluted in 3% BSA-PBS at a dilution of 1/100 overnight at 4 degree C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting

Immunohistochemistry analysis of GATA1 showing staining in the nucleus of paraffin-embedded human brain tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with GATA1 polyclonal antibody diluted in 3% BSA-PBS at a dilution of 1/100 overnight at 4 degree C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting

Immunohistochemistry analysis of GATA1 showing staining in the nucleus of paraffin-embedded human tonsil tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with GATA1 polyclonal antibody diluted in 3% BSA-PBS at a dilution of 1/100 overnight at 4 degree C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting

Western blot analysis of GATA1 was performed by loading 25 ug of various whole cell lysate onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. Membranes were then probed with rabbit polyclonal antibody recognizing GATA1 at a dilution of 1/500 overnight at 4 degree C on a rocking platform. Membranes were then washed in TBS-0.1% Tween 20 and probed with goat anti-rabbit-HRP secondary antibody for at least one hour. Membranes were washed and detection was carried out using chemiluminescence