PU.1/Spi-1 Antibody (823123)

Western blot shows lysates of NR8383 rat alveolar macrophage cell line and J774A.1 mouse reticulum cell sarcoma macrophage cell line. PVDF membrane was probed with 0.1 µg/mL of Rat Anti-Mouse PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB7124) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). 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.

PU.1/Spi-1 was detected in immersion fixed J774A.1 mouse reticulum cell sarcoma macrophage cell line using Rat Anti-Mouse PU.1/Spi-1 Monoclonal Antibody (Catalog # MAB7124) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red, upper panel; Catalog # NL013) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Mu
• Applications: WB, ICC/IF
• Clone: 823123
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Concentration: LYOPH

PU.1/Spi-1 Antibody (823123) Summary

• Immunogen: E. coli-derived recombinant mouse Spi-1
  Met1-Lys169
  Accession # P17433
• Specificity: Detects mouse PU.1/Spi-1 in ELISAs. In direct ELISAs, no cross-reactivity with recombinant human PU.1/Spi-1 is observed. Detects mouse and rat PU.1/Spi-1 in Western Blots
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat
• Gene: Spi1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for PU.1/Spi-1 Antibody (823123)

• 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

Background

Spi-1 (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). Spi-1 can act as both a transcriptional activator and repressor. In DC, Spi-1 promotes the expression of CD80, CD86 and CD11b, while in proerythroblasts it blocks GATA-1 induced transcriptional activation. Spi-1 binds to DNA as a monomer and interacts with multiple factors such as Runx-1, IRF8, GATA-1 and c-Jun. Mouse Spi-1 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 Spi-1 shares 88% and 81% aa identity with rat and human Spi-1, respectively.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PU.1/Spi-1 Antibody (MAB7124)(5)

Publications using MAB7124

• Wang J, Rattner A, Nathans J Bacterial meningitis in the early postnatal mouse studied at single-cell resolution eLife 2023-06-15 [PMID: 37318981]
• Roy ER, Chiu G, Li S et al. Concerted type I interferon signaling in microglia and neural cells promotes memory impairment associated with amyloid ? plaques Immunity 2022-05-01 [PMID: 35443157]
• J Wang, A Rattner, J Nathans A transcriptome atlas of the mouse iris at single-cell resolution defines cell types and the genomic response to pupil dilation Elife, 2021-11-16;10(0):. 11/16/2021 [PMID: 34783308]
• Neil Dani, Rebecca H. Herbst, Cristin McCabe, Gilad S. Green, Karol Kaiser, Joshua P. Head et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages Cell 5/1/2021 [PMID: 33932339]
• Neil Dani, Rebecca H. Herbst, Cristin McCabe, Gilad S. Green, Karol Kaiser, Joshua P. Head et al. A cellular and spatial map of the choroid plexus across brain ventricles and ages Cell 2021-05-01 [PMID: 33932339] (Immunohistochemistry, Mouse)

Bioinformatics

• Gene Symbol: Spi1
• Uniprot:
  • P17433(Mouse)