GATA-3 Antibody (634913)

R&D Systems | Catalog # MAB6330

R&D Systems

Key Product Details

Validated by

Orthogonal Validation

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Multiplex Immunofluorescence, Immunohistochemistry, Western Blot, Dual RNAscope ISH-IHC Compatible, Immunocytochemistry, Simple Western, COMET

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 634913
View all GATA-3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human GATA-3
Pro135-Ser258
Accession # P23771

Specificity

Detects human and mouse GATA-3 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for GATA-3 Antibody (634913)

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 antibody by Western Blot.

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 antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC).

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 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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 antibody by Simple WesternTM.

Detection of Human GATA‑3 by Simple WesternTM.

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 antibody in Human Breast Cancer Tissue Using Dual RNAscope® ISH and IHC.

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).
Detection of GATA-3 by Immunocytochemistry/ Immunofluorescence

Detection of GATA-3 by Immunocytochemistry/ Immunofluorescence

(A) qPCR was performed on groups of ten blastocysts to measure gene expression (n = 4–5; median ± IQR; ** p<0.01 Control vs. TMG; Mann–Whitney test). (B) Fixed blastocysts were stained with phalloidin (red), DAPI (blue) and anti-CDX2 antibody (green) or (C) anti-GATA3 antibody (green). Fluorescence microscopy was performed with optical sectioning, with maximum intensity projections shown. Scale bars 20 µm. (D) Quantification of anti-CDX2 and (E) anti-GATA3 antibody mean fluorescence intensity in blastocysts (arbitrary units). (n = 7–8 blastocysts from two independent experiments; median ± IQR; * p < 0.05 ** p < 0.01 Control vs. TMG; Mann–Whitney). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33036308), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GATA-3 by Immunohistochemistry

Detection of GATA-3 by Immunohistochemistry

Construction and characterization of kidney organoids. (A) Schematic diagram showing the stepwise induction of kidney organoids from H9 ESCs. (B) Morphology of H9 cells before organoid induction (day 0), H9-derived embryoid bodies, and cell assemblies/organoids at days 8 and 14 after induction, respectively. (C) Representative colonies of passage-30 H9 cells with immunostaining for the pluripotency-associated markers TRA-1-81 and OCT4 (scale bar = 20 μm). (D) H&E staining showing tubular profiles and multiple morphologies of cell assemblies within organoids sectioned at day 14. (E) Immunostaining of renal cell markers CDH1, MEIS1/2/3, LTL, NPHS1, LRP2, GATA3, UMOD, and CD31 (Scale bar = 20 μm, ×40 oil objective). (F) Temporal expressions of SLC12A1, NPSH1, GATA3, LRP2, and CD31 analyzed by qRT-PCR at days 0, 3, 8, and 14 (n = 3, *p < 0.05, ***p < 0.001). (G, H) Transmission electron microscopy showing podocyte-like cells (p) and proximal tubular epithelium-like cells (te) within the organoids at day 14. Note the foot processes (fp) anchored to the basement membrane-like profile (bm) and the oriented brush-like border (bb) (G: Scale = 2 μm; (H) Scale = 500 nm). (I) Schematic representation of kidney organoids (day 14). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38269090), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for GATA-3 Antibody (634913)

Application
Recommended Usage

COMET

Optimal dilutions of this antibody should be experimentally determined.

Dual RNAscope ISH-IHC Compatible

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed MCF-7 human breast cancer cell line

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue

Multiplex Immunofluorescence

20 µg/mL
Sample: Paraffin embedded tissue sections of Human Breast Cancer

Simple Western

10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line and MCF‑7 human breast cancer cell line

Western Blot

0.1 µg/mL
Sample: 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

Reviewed Applications

Read 1 review rated 4 using MAB6330 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GATA-3

GATA-3 belongs to the GATA family of transcription factors, which bind to the consensus DNA sequence (A/T) GATA (A/G) to control diverse tissue-specific programs of gene expression and morphogenesis. It is widely expressed in mesodermal- and endodermal-derived tissues. GATA-3 has been shown to be an essential regulator for immune cell function, sympathetic neuron development and the maintenance of the differentiated state in epithelial cells.

Alternate Names

GATA3

Entrez Gene IDs

2625 (Human); 14462 (Mouse)

Gene Symbol

GATA3

UniProt

P23771

Additional GATA-3 Products

Product Documents for GATA-3 Antibody (634913)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for GATA-3 Antibody (634913)

For research use only

Citations for GATA-3 Antibody (634913)

Customer Reviews for GATA-3 Antibody (634913) (1)

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  • Human/Mouse GATA-3 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: BeWo human choriocarcinoma cell line
    Species: Human
    Verified Customer | Posted 03/20/2023
    BeWo Cell. Gata3 & Merge with DAPI.
    GATA-3 Antibody (634913) MAB6330

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