Detects mouse PU.1/Spi-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 30% cross-reactivity with recombinant human Spi-1 is observed, and less than 1% cross-reactivity with recombinant mouse Spi-B is observed.
Polyclonal Sheep IgG
E. coli-derived recombinant mouse PU.1/Spi-1 Met1-Lys169 Accession # P17433
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.
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 Mouse PU.1/Spi‑1 by Western Blot. Western blot shows lysates of J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Mouse PU.1/Spi‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7124) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). 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.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 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.
Spi1 (SFFV proviral integration 1 protein; also PU.1 and Sfpi1) is a 37-41 kDa member of the ets family of transcription factors. It is both an RNA and DNA binding protein that is found in hematopoietic cells such as B cells, neutrophils, macrophages and dendritic cells (DC). Spi1 can act as both a transcriptional activator and repressor. In DC, Spi1 promotes the expression of CD80, CD86 and CD11b, while in proerythroblasts, it blocks GATA-1 induced transcriptional activation. Spi1 binds to DNA as a monomer, and interacts with multiple factors such as Runx-1, IRF8, GATA-1 and c-Jun. Mouse Spi1 is 272 amino acids (aa) in length. It contains a transactivation region (aa 7-117), a PEST domain (aa 118-164), and a DNA-binding domain (aa 165-266). Phosphorylation on Ser41, 142 and 148 increases activity. There is one alternative start site at Met7. Over aa 1-169, mouse Spi1 shares 88% and 81% aa identity with rat and human Spi1, respectively.
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