Rabbit anti-Human GATA3 Polyclonal Antibody | anti-GATA3 antibody
Rabbit anti GATA3
IF: 1:50-1:500
WB: 1:1000
IHC-Paraffin: 1:50-1:500
Storage in frost-free freezers is not recommended. This product should be stored undiluted.
Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. Shelf Life: 12 months from date of despatch
Western Blot (WB)
(Western blot analysis of GATA3 was performed by loading 15 ug of SH-SY5Y whole cell lysate (WCL), SH-SY5Y nuclear extract (NE) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked for at least 1 hour. The membrane was probed with GATA3 polyclonal antibody at a dilution of 1/1000 overnight at 4 degree C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody for 1 hour. Chemiluminescence was performed for band detection)
Immunohistochemistry (IHC)
(Immunohistochemistry analysis of GATA3 showing staining in the nucleus of paraffin-embedded human tonsil tissue (right) compared to 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 GATA3 polyclonal antibody diluted in 3% BSA-PBS at a dilution of 1/200 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)
Immunofluorescence (IF)
(Immunofluorescent analysis of GATA3 (green) in MCF7 (left panel) and SH-SY5Y cells (right panel). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with GATA3 polyclonal antibody at a dilution of 1/50 for at least 1 hour at room temperature, washed with PBS, and incubated with a goat anti-rabbit IgG secondary antibody (green) for 30 minutes at room temperature. Cells were counterstained with phalloidin (F-actin, red) and Hoechst 33342 (Nucleus, blue). Cells were viewed at 20x magnification)
GATA3 has been shown to play a critical role in T cell development and the differentiation of T helper 2 (Th2) cells (Ho et al.1991). In addition, GATA3 has been implied to have tissue specific functions such as a regulatory role in the development of the mammary gland epithelium and skin tissue (Chou et al. 2010).
Aberrant GATA3 levels have been identified in a variety of cancers including breast cancer. Indeed, GATA3 has become a marker for certain breast cancer types and GATA3 antibodies are commonly used for the immunohistochemical evaluation of breast cancer samples (Krings et al. 2014).
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a protein which belongs to the GATA family of transcription factors. The protein contains two GATA-type zinc fingers and is an important regulator of T-cell development and plays an important role in endothelial cell biology. Defects in this gene are the cause of hypoparathyroidism with sensorineural deafness and renal dysplasia. [provided by RefSeq, Nov 2009]
Uniprot Description
Transcriptional activator which binds to the enhancer of the T-cell receptor alpha and delta genes. Binds to the consensus sequence 5'-AGATAG-3'. Required for the T-helper 2 (Th2) differentiation process following immune and inflammatory responses.
