Mouse PU.1/Spi-1 Antibody

  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse PU.1/Spi-1 in ELISAs. In direct ELISAs, no cross-reactivity with recombinant human PU.1/Spi-1 is observed. Detects mouse and rat PU.1/Spi-1 in Western Blots
  • Source
    Monoclonal Rat IgG2A Clone # 823123
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant mouse Spi-1
    Met1-Lys169
    Accession # P17433
  • 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
    0.1 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    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 Mouse and Rat PU.1/Spi‑1 by Western Blot. Western blot shows lysates of NR8383 rat alveolar macrophage cell line and J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Mouse PU.1/Spi‑1 Monoclonal Antibody (Catalog # MAB7124) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for PU.1/Spi‑1 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
PU.1/Spi‑1 in J774A.1 Mouse Cell Line. PU.1/Spi‑1 was detected in immersion fixed J774A.1 mouse reticulum cell sarcoma macrophage cell line using Rat Anti-Mouse PU.1/Spi‑1 Monoclonal Antibody (Catalog # MAB7124) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red, upper panel; Catalog # NL013) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
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: PU.1/Spi-1

Spi-1 (SFFV proviral integration 1 protein; also PU.1 and Sfpi1) is a 37-41 kDa member of the ets family of transcription factors. It is both an RNA and DNA binding protein that is found in hematopoietic cells such as B cells, neutrophils, macrophages and dendritic cells (DC). Spi-1 can act as both a transcriptional activator and repressor. In DC, Spi-1 promotes the expression of CD80, CD86 and CD11b, while in proerythroblasts it blocks GATA-1 induced transcriptional activation. Spi-1 binds to DNA as a monomer and interacts with multiple factors such as Runx-1, IRF8, GATA-1 and c-Jun. Mouse Spi-1 is 272 amino acids (aa) in length. It contains a transactivation region (aa 7-117), a PEST domain (aa 118-164), and a DNA-binding domain (aa 165-266). Phosphorylation on Ser41, 142 and 148 increases activity. There is one alternative start site at Met7. Over aa 1-169, mouse Spi-1 shares 88% and 81% aa identity with rat and human Spi-1, respectively.

  • 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
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