Immunofluorescence analysis of GATA3 was done on 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with GATA3 Rabbit Polyclonal antibody (PA1101) at 1ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified partial recombinant GATA3 protein expressed in E. coli|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||1:500-1:1500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Western blot analysis of PA1-101 detects a prominent ~50 kDa protein in GATA3 expressing SH-SY5Y cells. The signal is enhanced in nuclear extracts due to enrichment of GATA3. This antibody has also been validated for Immunofluorescence and IHC (P).PA1-101 was successfully used in western blotting analysis of GATA3 in breast cancer cell lines.
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. GATA3 also regulates luminal epithelial cell differentiation in the mammary gland and is mutated in breast cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.