Human HNF-4 alpha /NR2A1 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human HNF‑4 alpha /NR2A1 in ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG2B Clone # 843716
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human HNF‑4 alpha /NR2A1
    Val130-Ser330
    Accession # P41235
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human HNF‑4 alpha /NR2A1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. Gels were loaded with 10 µg of cytoplasmic (Cyto) and 5 µg of nuclear (Nuc) extracts. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human HNF‑4 alpha /
NR2A1 Monoclonal Antibody (Catalog # MAB4605) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). Specific bands were detected for HNF‑4 alpha /NR2A1 at approximately 50-55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
HNF-4 alpha /NR2A1 in HepG2 Human Cell Line. HNF‑4 alpha /NR2A1 was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Mouse Anti-Human HNF‑4 alpha /NR2A1 Monoclonal Antibody (Catalog # MAB4605) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human HNF‑4 alpha /NR2A1 by Simple WesternTM.

Simple Western lane view shows lysates of human liver tissue, loaded at 0.2 mg/mL. A specific band was detected for HNF‑4 alpha /NR2A1 at approximately 62 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human HNF‑4 alpha /NR2A1 Monoclonal Antibody (Catalog # MAB4605). 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.

Intracellular Staining by Flow Cytometry
Detection of HNF-4 alpha/NR2A1 in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Mouse Anti-Human HNF-4 alpha/NR2A1 Monoclonal Antibody (Catalog # MAB4605, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram) followed by PE-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of HNF-4 alpha/NR2A1 in Human Endodermal Cells by Flow Cytometry. BG01V human embryonic stem cell line differentiated to endodermal cells (StemXVivo Endoderm Kit, Catalog # SC019B) was stained with Mouse Anti-Human HNF-4 alpha/NR2A1 Monoclonal Antibody (Catalog # MAB4605, filled histogram or isotype control antibody (Catalog # MAB0041, open histogram) followed by PE-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.                               
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    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.
Background: HNF-4 alpha/NR2A1
HNF-4 alpha is a transcription factor that binds DNA as a homodimer. HNF‑4 alpha is important in liver, kidney, and intestinal development. It has also been intensely studied as one of a variety of genes responsible for diabetes mellitus. HNF-4 alpha has been shown in knock out mice to be essential for the morphogenic and functional differentiation of hepatocytes. HNF-4 alpha is a dominant regulator of epithelial phenotypes able to drive the mesenchymal-to-epithelial transition when expressed in fibroblasts.
  • Long Name:
    Hepatocyte Nuclear Factor-4, alpha
  • Entrez Gene IDs:
    3172 (Human)
  • Alternate Names:
    FLJ39654; hepatic nuclear factor 4 alpha; hepatocyte nuclear factor 4, alpha; hepatocyte nuclear factor 4-alpha; HNF4 alpha; HNF-4 alpha; HNF4A; HNF4a7; HNF-4-alpha; HNF4alpha10/11/12; HNF4HNF4a8; MODY; MODY1; NR2A1; NR2A1HNF4a9; NR2A21; Nuclear receptor subfamily 2 group A member 1; TCF; TCF14; TCF-14; TCF14HNF4alpha; Transcription factor 14; Transcription factor HNF-4; transcription factor-14
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