Human PU.1/Spi-1 PE-conjugated Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human PU.1/Spi‑1 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human SPI-B is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human PU.1/Spi‑1
    Met1-Lys169
    Accession # NP_001074016
  • Formulation
    Supplied in a saline solution containing BSA and Sodium Azide.
  • Label
    Phycoerythrin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    10 µL/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
Intracellular Staining by Flow Cytometry

Detection of PU.1/Spi‑1 in Human PBMC lymphocytes by Flow Cytometry. Human peripheral blood mononuclear cell (PBMC) lymphocytes were stained with Sheep Anti-Human PU.1/Spi‑1 PE‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # IC5870P) and Mouse Anti-Human CD3 epsilon PerCP‑conjugated Monoclonal Antibody (Catalog # FAB100C). Quadrant markers were set based on control antibody staining (Catalog # IC016P). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Intracellular Staining by Flow Cytometry

Detection of PU.1/Spi‑1 in Th2-differentiated Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with 5 ng/mL Recombinant Human IL‑4 (Catalog # 204-IL) and 10 μg/mL Goat Anti-Human IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-285-NA) for 5 days, then treated overnight with PMA and Calcium Ionomycin to induce Th2 differentiation were stained with Sheep Anti-Human PU.1/Spi‑1 PE‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # IC5870P) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # IC016P). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: PU.1/Spi-1
PU.1 (Purine-rich Nucleic Acid Binding Protein 1), also known as 31 kDa Transforming Protein and SPI-1, is a member of the PU subfamily, ETS family of transcription factors. Although its predicted MW is 31 kDa, it appears to run anomalously high in SDS-PAGE at 40-45 kDa. PU.1 is a monomeric hematopoietic protein that regulates the differentiation of early myeloid and lymphoid progenitors. High PU.1 levels favor granulocyte and macrophage production, while low levels generate megakaryocytes, erythrocytes, T and B cells. Human PU.1 is 270 amino acids (aa) in length. It contains an N-terminal acidic/polyGln transactivation region (aa 34‑99), a non-destabilizing PEST sequence (aa 117-165) and a C-terminal Ets DNA-binding domain (aa 170-253). PU.1 is phosphorylated on Ser146, allowing it to bind to Pip. Over aa 1-169, human PU.1 shares 88% aa identity with mouse PU.1.
  • Long Name:
    Hematopoietic transcription factor PU.1
  • Entrez Gene IDs:
    6688 (Human); 20375 (Mouse); 366126 (Rat)
  • Alternate Names:
    hematopoietic transcription factor PU.1; OF; PU.1; SFPI1; SFPI131 kDa transforming protein; SPI-1 proto-oncogene; SPI1; Spi-1; SPI-A; spleen focus forming virus (SFFV) proviral integration oncogene spi1,31 kDa-transforming protein; transcription factor PU.1
Related Research Areas
Isotype Controls
Description Application Cat# Citations Images  

Sheep IgG PE-conjugated Antibody

Ctrl IC016P  
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Staining Reagents
Description Application Cat# Citations Images  

Flow Cytometry Permeabilization/Wash Buffer I (1X)

Flow FC005 5
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Flow Cytometry Fixation Buffer (1X)

Flow FC004 3
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Flow Cytometry Fixation & Permeabilization Buffer Kit I

Flow FC009 3
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Flow Cytometry Fixation/Permeabilization Buffer I (1X)

Flow FC007 2
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