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

Name: Human PU.1/Spi‑1 PE‑conjugated Antibody
Brand: R&D Systems
SKU: IC5870P

R&D Systems | Catalog # IC5870P

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Product Documents

Discontinued Product

IC5870P has been discontinued. View all PU.1/Spi-1 products.

Product Specifications
Scientific Data
Applications
Spectra Viewer
Flow Cytometry
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Intracellular Staining by Flow Cytometry

Cited:

Flow Cytometry

Label

Phycoerythrin (Excitation = 488 nm, Emission = 565-605 nm)

Antibody Source

Polyclonal Sheep IgG

View all PU.1/Spi-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human PU.1/Spi‑1
Met1-Lys169
Accession # NP_001074016

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human PU.1/Spi‑1 PE‑conjugated Antibody

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 CD3e 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.

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 CD3e 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.

Applications for Human PU.1/Spi‑1 PE‑conjugated Antibody

Application

Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/10⁶ cells
Sample: Human peripheral blood mononuclear cell (PBMC) lymphocytes and Human peripheral blood mononuclear cells (PBMCs) treated with Recombinant Human IL‑4 (Catalog # 204-IL) and 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 fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005).

Spectra Viewer

Plan Your Experiments

Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

Spectra Viewer

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
Spillover Popups - Visualize the spectra of individual fluorochromes
Antigen Density Selector - Match fluorochrome brightness with antigen density

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Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.

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.