Human PU.1/Spi-1 Antibody

Catalog # Availability Size / Price Qty
MAB5870
MAB5870-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/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.
Source
Monoclonal Mouse IgG2B Clone # 732322
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant human PU.1/Spi-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
2 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.
 
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
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

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

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.

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

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.

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
Sterile PBS to a final concentration of 0.5 mg/mL.
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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Isotype Controls

Reconstitution Buffers

Secondary Antibodies

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