Human PU.1/Spi-1 Antibody

Catalog #: AF5870 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF5870
AF5870-SP

Detection of Human PU.1 by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

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 either lyophilized or 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.

Scientific Data

Western Blot

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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

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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

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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