Human/Mouse GATA-3 Antibody

Detection of GATA-3 in Human Breast Cancer via Multiplex Immunofluorescence staining on COMET™​ GATA-3 was detected in immersion fixed paraffin-embedded sections of human breast cancer using Mouse Anti-Human GATA‑3 Monoclonal Antibody (MAB6330) at 20 µg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the nucleus with dim cytoplasmic background signal. Protocol available in COMET™ Panel Builder.

Detection of Human and Mouse GATA‑3 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, MCF-7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, and EL-4 mouse lymphoblast cell line. PVDF Membrane was probed with 0.1 µg/mL of Moue Anti-Human/Mouse GATA-3 Monoclonal Antibody (Catalog # MAB6330) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). Specific bands were detected for full length (FL) GATA-3 at approximately 52 kDa and the splice form (SF) found in MCF-7 cells at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

GATA‑3 in MCF‑7 Human Cell Line. GATA-3 was detected in immersion fixed MCF-7 human breast cancer cell line using Mouse Anti-Human/Mouse GATA-3 Monoclonal Antibody (Catalog # MAB6330) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

GATA‑3 in Human Breast Cancer Tissue. GATA-3 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human/Mouse GATA-3 Monoclonal Antibody (Catalog # MAB6330) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human GATA‑3 by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line, loaded at 0.5 mg/mL. Specific bands were detected for GATA‑3 at approximately 53 (splice variant) and 61 kDa (full length) as indicated, using 10 µg/mL of Mouse Anti-Human/Mouse GATA‑3 Monoclonal Antibody (Catalog # MAB6330). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

GATA-3 in Human Breast Cancer Tissue Using Dual RNAscope® ISH and IHC. GATA-3 mRNA (red) and protein (green) was detected in formalin-fixed paraffin-embedded tissue sections of human breast cancer tissue probed with ACD RNAScope® Probe (Catalog # 403551) followed by immunohistochemistry using R&D Systems Mouse Anti-Human/Mouse GATA-3 Monoclonal Antibody (Catalog# MAB6330) at 5 μg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte HRP Polymer Antibody (VC001). Tissue was stained using ACD RNAscope® 2.5 HD Duplex Detection Reagents (Catalog # 322500).