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_001074016Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Phycoerythrin
Applications
Recommended
ConcentrationSample
Intracellular Staining by Flow Cytometry
10 µL/10 6 cells
See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. 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 # ). Quadrant markers were set based on control antibody staining (Catalog # ). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # ) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # ). View our protocol for . |
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 # ) and 10 μg/mL Goat Anti-Human IFN‑ gamma Antigen Affinity-purified Polyclonal Antibody (Catalog # ) 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 # ). Quadrant markers were set based on control antibody staining (Catalog # ). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # ) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # ). View our protocol for . |
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
