Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Rat, Chimpanzee, Primate - Macaca mulatta (Rhesus Macaque), Transgenic Rat, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all HNF-3 beta/FoxA2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human HNF‑3 beta /FoxA2
Met242-Ser457
Accession # Q9Y261

Specificity

Detects human HNF‑3 beta /FoxA2 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human HNF‑3 beta /FoxA2 Antibody

Detection of HNF-3 beta /FoxA2-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of HNF‑3 beta /FoxA2-regulated Genes by Chromatin Immunoprecipitation.

Mouse splenocytes were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. HNF-3 beta /FoxA2/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human HNF-3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. TheE-RABPpromoter was detected by standard PCR.
HNF-3 beta /FoxA2 antibody in Endoderm Differentiated BG01V Human Stem Cells by Immunocytochemistry (ICC).

HNF‑3 beta /FoxA2 in Endoderm Differentiated BG01V Human Stem Cells.

HNF-3 beta /FoxA2 was detected in immersion fixed endoderm differentiated BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human HNF-3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
HNF-3 beta /FoxA2 antibody in Human Liver by Immunohistochemistry (IHC-P).

HNF‑3 beta /FoxA2 in Human Liver.

HNF-3 beta /FoxA2 was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human HNF-3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of HNF‑3 beta /FoxA2 in HepG2 Cells (Positive) and THP‑1 Cells (Negative).

HNF‑3 beta /FoxA2 was detected in immersion fixed HepG2 Human Hepatocellular Carcinoma Cells (Positive) and absent in THP‑1 Human Acute Monocytic Leukemia Cells (Negative) using Goat Anti-Human HNF‑3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) at 5 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 cell nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human HNF‑3 beta /FoxA2 by Western Blot.

Western blot shows lysates of HepG2 human hepatocellular carcinoma cells. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human HNF‑3 beta /FoxA2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for HNF‑3 beta /FoxA2 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Detection of Human HNF-3 beta/FoxA2 by Immunocytochemistry/Immunofluorescence

Detection of Human HNF-3 beta/FoxA2 by Immunocytochemistry/Immunofluorescence

Differentiation of hESCs into definitive endoderm cells by Activin A with CHIR99021.a The relative primitive streak (BRACHYRUY, MESP1, and MIXL1) gene expression of the day 1 differentiated cells by adding Activin A (100 ng/mL) with CHIR99021 (2–5 µM) or Wnt3a (25–100 ng/mL) treatments were determined by real-time quantitative PCR (qPCR). None, no WNT signaling pathway activators were used on day 1 for PS differentiation (Activin A only). b, c After 1 day, the medium was changed to Activin A (100 ng/mL) and 1 × CTS-B27 to induce DE differentiation. The definitive endoderm (FOXA2 and SOX17) and mesoderm (MESP2 and HAND1) relative gene expression levels were determined by qPCR. d Immunofluorescence analysis of the expression of SOX17 and FoxA2 for Activin A with 2 µM CHIR99021-induced differentiated cells on day 3. e, f The expression of SOX17 and FoxA2 for Activin A with 2 µM CHIR99021-induced differentiated cells was determined by flow cytometry on day 3. Isotype control antibodies were used as controls. At a specific gene expression, datum points lacking common letters differ, p < 0.05. Data are represented as the mean ± SD. Scale bar, 100 µm Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31601782), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunofluorescent Staining of Adult Stem Cell-derived Liver Organoids.

Adult stem cell-derived liver organoids were generated following the steps detailed in the human liver organoid culture protocol. Differentiated human liver organoids were stained using a (A) Mouse Anti-Human Serum Albumin Monoclonal Antibody (Catalog # MAB1455; red) and a (B) Goat Anti-Human HNF-3beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2400; red) and counterstained with DAPI (Catalog # 5748; blue).
Detection of HNF-3 beta /FoxA2 by Flow Cytometry

Detection of HNF-3 beta /FoxA2 by Flow Cytometry

FOXA2 drives the p3-specific chromatin accessibility program(A) Footprinting score for the FOX motif is highest in p3 samples.(B) Foxa2 expression in p3 NPs suggests it is the most likely candidate to drive the footprinting signal.(C) Average ATAC-seq accessibility at FOXA2 ChIP-seq peaks13 in the indicated samples shows these regions are highly accessible in p3 NPs.(D) Normalized FOXA2 ChIP-seq coverage showing accessibility in p3-specific elements from the groups of ATAC-seq elements in the indicated clusters from Figure 3B.(E) Genetic lineage tracing indicates that cells that expressed Foxa2 at E8.5 (tamoxifen administration) have contributed to the p3 progenitor and V3 neuronal cell types by E11.5 (red arrows).(F) Quantifications of p3 and V3 cells expressing tdTomato in embryos collected after tamoxifen administration at the indicated times. Biological replicates: n = 2 for E7.5, n = 4 for E8.5, n = 3 for E9.5.(G) Foxa2−/− ES cells fail to generate p3 NPs when exposed to 500 nM SAG.(H) Representative flow cytometry plots of the quantifications in (G) showing a marked reduction in p3 NPs from Foxa2−/− ES cells compared with wild type. Cells are gated for SOX2+ live neural progenitors.(I) Representative immunohistochemistry staining for SOX2, OLIG2 and NKX2,2 showing reduced number of cells expressing NKX2.2 in Foxa2−/− mutant cells at day 6 of differentiation treated with 500 nM SAG.Scale bars, 50 μm.See also Figure S4. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36516856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HNF-3 beta /FoxA2 by Immunocytochemistry/ Immunofluorescence

Detection of HNF-3 beta /FoxA2 by Immunocytochemistry/ Immunofluorescence

Expression profiles and functional enrichments of marker genes across time points identified during hESC differentiation. A. UMAP visualization of the expression of well-defined marker genes of different stages in hESC differentiation. B. The representative immunofluorescence staining for definitive endoderm cells with antibodies against SOX17 and FOXA2. DAPI serves as a nucleus indicator. The individual color channels were merged to assess the colocalization of SOX17 an FOXA2 expression in the nuclei. Scale bars, 200 μm. C. Dot plot showing the average and percentage expression of well-defined marker genes in different stages or cell types. Genes are colored according to their mean expression level. Diameter denotes fractional expression. SC-alpha, SC-beta, and SC-EC represent stem-cell-derived alpha, beta, and enterochromaffin cells, respectively. D. Heatmap showing the expression of the top ten marker genes of each time point. lncRNA genes are indicated by arrows. E. Enriched GO terms of marker genes of each time point. F. UMAP visualization of the expression of genes related to endoderm formation. G. Violin plots showing the expression of lncRNA genes that listed in the top 10 marker genes of each time point Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36907881), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HNF-3 beta /FoxA2 by Immunocytochemistry/ Immunofluorescence

Detection of HNF-3 beta /FoxA2 by Immunocytochemistry/ Immunofluorescence

Expression profiles and functional enrichments of marker genes across time points identified during hESC differentiation. A. UMAP visualization of the expression of well-defined marker genes of different stages in hESC differentiation. B. The representative immunofluorescence staining for definitive endoderm cells with antibodies against SOX17 and FOXA2. DAPI serves as a nucleus indicator. The individual color channels were merged to assess the colocalization of SOX17 an FOXA2 expression in the nuclei. Scale bars, 200 μm. C. Dot plot showing the average and percentage expression of well-defined marker genes in different stages or cell types. Genes are colored according to their mean expression level. Diameter denotes fractional expression. SC-alpha, SC-beta, and SC-EC represent stem-cell-derived alpha, beta, and enterochromaffin cells, respectively. D. Heatmap showing the expression of the top ten marker genes of each time point. lncRNA genes are indicated by arrows. E. Enriched GO terms of marker genes of each time point. F. UMAP visualization of the expression of genes related to endoderm formation. G. Violin plots showing the expression of lncRNA genes that listed in the top 10 marker genes of each time point Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36907881), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HNF-3 beta /FoxA2 by Flow Cytometry

Detection of HNF-3 beta /FoxA2 by Flow Cytometry

FOXA2 drives the p3-specific chromatin accessibility program(A) Footprinting score for the FOX motif is highest in p3 samples.(B) Foxa2 expression in p3 NPs suggests it is the most likely candidate to drive the footprinting signal.(C) Average ATAC-seq accessibility at FOXA2 ChIP-seq peaks13 in the indicated samples shows these regions are highly accessible in p3 NPs.(D) Normalized FOXA2 ChIP-seq coverage showing accessibility in p3-specific elements from the groups of ATAC-seq elements in the indicated clusters from Figure 3B.(E) Genetic lineage tracing indicates that cells that expressed Foxa2 at E8.5 (tamoxifen administration) have contributed to the p3 progenitor and V3 neuronal cell types by E11.5 (red arrows).(F) Quantifications of p3 and V3 cells expressing tdTomato in embryos collected after tamoxifen administration at the indicated times. Biological replicates: n = 2 for E7.5, n = 4 for E8.5, n = 3 for E9.5.(G) Foxa2−/− ES cells fail to generate p3 NPs when exposed to 500 nM SAG.(H) Representative flow cytometry plots of the quantifications in (G) showing a marked reduction in p3 NPs from Foxa2−/− ES cells compared with wild type. Cells are gated for SOX2+ live neural progenitors.(I) Representative immunohistochemistry staining for SOX2, OLIG2 and NKX2,2 showing reduced number of cells expressing NKX2.2 in Foxa2−/− mutant cells at day 6 of differentiation treated with 500 nM SAG.Scale bars, 50 μm.See also Figure S4. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36516856), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human HNF‑3 beta /FoxA2 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: Mouse splenocyte chromatin, E-RABP promoter detected by standard PCR.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed endoderm differentiated BG01V human embryonic stem cells and immersion fixed HepG2 Human Hepatocellular Carcinoma Cells (Positive) and absent in THP‑1 Human Acute Monocytic Leukemia Cells (Negative)

Immunohistochemistry

3-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human liver

Western Blot

1 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line

Reviewed Applications

Read 3 reviews rated 4.7 using AF2400 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 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. *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: HNF-3 beta/FoxA2

HNF-3 beta, also known as FoxA2, is a member of the forkhead class of DNA-binding proteins. It is a transcriptional activator for liver-specific transcripts such as albumin and transthyretin. Similar family members play roles in the differentiation of the pancreas and liver.

Long Name

Hepatocyte Nuclear Factor-3, beta

Alternate Names

FoxA2, HNF3 beta

Entrez Gene IDs

3170 (Human)

Gene Symbol

FOXA2

UniProt

Q9Y261

Additional HNF-3 beta/FoxA2 Products

Product Documents for Human HNF‑3 beta /FoxA2 Antibody

Certificate of Analysis

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Product Specific Notices for Human HNF‑3 beta /FoxA2 Antibody

For research use only

Citations for Human HNF‑3 beta /FoxA2 Antibody

Customer Reviews for Human HNF‑3 beta /FoxA2 Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
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4 Stars
33%
3 Stars
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Showing  1 - 3 of 3 reviews Showing All
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  • Good product!
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Midbrain organoid derived from iPSC
    Species: Midbrain organoid derived from iPSC
    Verified Customer | Posted 01/21/2026
    FOXA2-488 OTX2-568 DAPI
    Works well on our tissue. Dilution used: 1:100
    Human HNF‑3 beta /FoxA2 Antibody AF2400
  • Human HNF-3 beta /FoxA2 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Prostate cancer tissue
    Species: Human
    Verified Customer | Posted 09/10/2021
    Human HNF‑3 beta /FoxA2 Antibody AF2400
  • Human HNF-3 beta /FoxA2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: alveoli
    Species: Human
    Verified Customer | Posted 02/15/2019
    dapi- BLUE magenta - foxa2
    Human HNF‑3 beta /FoxA2 Antibody AF2400

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