Detection of Human GATA‑5 by Western Blot.
Western blot shows lysates of HEK293 human embryonic kidney cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human GATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for GATA‑5 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of GATA‑5-regulated Genes by Chromatin Immunoprecipitation.
HeLa human cervical epithelial carcinoma cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. GATA‑5/DNA complexes were immunoprecipitated using 5 μg Goat Anti-Human GATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The mucin4 promoter was detected by standard PCR.
GATA‑5 in HepG2 Human Cell Line.
GATA‑5 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Goat Anti-Human GATA‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2170) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
GATA-5 belongs to the GATA family of transcription factors which bind to the consensus DNA sequence (AIT) GATA (AIG) to control diverse tissue-specific programs of gene expression and morphogenesis. GATA5 is important in the development and differentiation of endoderm-derived organs, including heart and gastrointestinal tissues. GATA-5 has also been implicated in cancer development (1, 2).
Tsai, S.F. et al. (1989) Nature 339:446.
Jiang, Y. and T. Evans (1996) Dev. Biol. 174:258.
Entrez Gene IDs:
bB379O24.1; GATA binding factor-5; GATA binding protein 5; GATA5; GATA-5; GATA-binding factor 5; GATA-binding protein 5; transcription factor GATA-5
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