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.
Monoclonal Mouse IgG2B Clone # 732322
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human PU.1/Spi-1 Met1-Lys169 Accession # NP_001074016
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.
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
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.
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.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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.
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.
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