Human PU.1/Spi-1 Antibody

Catalog # Availability Size / Price Qty
AF5870
AF5870-SP
Human PU.1/Spi-1 Antibody in Western Blot
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Human PU.1/Spi-1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human PU.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
Met1-Lys169
Accession # NP_001074016
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.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
PMA and Ca2+ ionomycin‑treated human Th2-stimulated PBMCs, fixed with paraformaldehyde, and permeabilized with saponin

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

Western Blot Detection of Human PU.1 by Western Blot. View Larger

Detection of Human PU.1 by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line. PVDF membrane was probed with 1 µg/mL of Human PU.1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5870) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PU.1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunocytochemistry PU.1 in THP‑1 Human Cell Line. View Larger

PU.1 in THP‑1 Human Cell Line. PU.1 was detected in immersion fixed THP-1 human acute monocytic leukemia cell line using Human PU.1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5870) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Reconstitution Calculator

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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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

PU.1 (Purine-rich nucleic acid binding protein 1; also 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 magakaryocytes, 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

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