Overview

  • Product name
  • Description
    Mouse monoclonal to PU.1/Spi1
  • Tested applications
    Suitable for: WB, ELISA, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment, corresponding to amino acids 7-126 of human PU.1/Spi1 (NP_003111) with a proprietary tag.

  • Positive control
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab76542 as a replacement.

Properties

Applications

Our Abpromise guarantee covers the use of ab88082 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 31 kDa.
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.
Flow Cyt Use 2µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Binds to the PU-box, a purine-rich DNA sequence (5'-GAGGAA-3') that can act as a lymphoid-specific enhancer. This protein is a transcriptional activator that may be specifically involved in the differentiation or activation of macrophages or B-cells. Also binds RNA and may modulate pre-mRNA splicing.
  • Sequence similarities
    Belongs to the ETS family.
    Contains 1 ETS DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • transcription factor spi1 antibody
    • 31 kDa Transforming Protein antibody
    • 31 kDa-transforming protein antibody
    • cb1086 antibody
    • Hematopoietic transcription factor PU.1 antibody
    • OF antibody
    • oncogene spi1 antibody
    • PU.1 antibody
    • SFFV virus-induced murine erythroleukemia oncogene, mouse, homolog of antibody
    • SFPI1 antibody
    • si:by184l24.2 antibody
    • SPI 1 antibody
    • SPI 1 proto oncogene antibody
    • SPI A antibody
    • Spi1 antibody
    • SPI1_HUMAN antibody
    • Spleen focus forming virus (SFFV) proviral integration oncogene spi1 antibody
    • Spleen focus forming virus proviral integration oncogene spi1 antibody
    • Transcription factor PU.1 antibody
    see all

Images

  • ab88082, at 10 µg/ml, staining PU.1/Spi1 in HeLa cells by Immunofluorescence.
  • Detection limit for recombinant tagged ab88082 is approximately 0.1ng/ml as a capture antibody.
  • Anti-PU.1/Spi1 antibody (ab88082) at 5 µg/ml + tagged recombinant protein fragment (the immunogen) at 0.2 µg

    Secondary
    Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/5000 dilution

    Predicted band size : 31 kDa
    Observed band size : 39 kDa (why is the actual band size different from the predicted?)
    Note: Molecular weight of the proprietary tag alone is 26 kDa.
  • Anti-PU.1/Spi1 antibody (ab88082) at 5 µg/ml + Raw 264.7 cell lysate at 25 µg

    Secondary
    Goat anti-Mouse IgG (H&L)-HRP at 1/2500 dilution

    Predicted band size : 31 kDa
    Observed band size : 31 kDa
  • Ab88082 staining PU.1/Spi1 in human induced microglia by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde for 15 minutes. Samples were incubated with primary antibody (1:200) for 2 hours at room temperature. Hoechst nuclear stain (blue) was used at 1:5000 dilution. 

  • Overlay histogram showing K562 cells stained with ab88082 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab88082, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Sellgren CM  et al. Patient-specific models of microglia-mediated engulfment of synapses and neural progenitors. Mol Psychiatry 22:170-177 (2017). ICC/IF ; Human . Read more (PubMed: 27956744) »
  • Zhou J  et al. PU.1 is essential for MLL leukemia partially via crosstalk with the MEIS/HOX pathway. Leukemia 28:1436-48 (2014). Read more (PubMed: 24445817) »

See all 2 Publications for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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