Human PU.1/Spi‑1 Antibody

Name: Human PU.1/Spi‑1 Antibody
Brand: R&D Systems
SKU: MAB5870
Price: 459 USD
Availability: InStock

R&D Systems | Catalog # MAB5870

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

Key Product Details

Species Reactivity

Human

Applications

Western Blot, Intracellular Staining by Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2B Clone # 732322

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, no cross-reactivity with recombinant human (rh) Spi-B, recombinant mouse (rm) PU.1/Spi-1, or rmSpi-B is observed. In Western blots, approximately 25% cross-reactivity with rhSpi-B and rmPU.1/Spi-1 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2B

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

Detection of Human PU.1/Spi‑1 by Western Blot.

Western blot shows lysates of K562 human chronic myelogenous leukemia cell line and Daudi human Burkitt's lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB5870) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). 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.

Detection of PU.1/Spi‑1 in THP‑1 Human Cell Line by Flow Cytometry.

THP-1 human acute monocytic leukemia cell line was stained with Mouse Anti-Human PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB5870, filled histogram) or isotype control antibody (Catalog # MAB004, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Applications for Human PU.1/Spi‑1 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells
Sample: THP‑1 human acute monocytic leukemia cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

2 µg/mL
Sample: K562 human chronic myelogenous leukemia cell line and Daudi human Burkitt's lymphoma cell line

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

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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.