Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

PU.1 (9G7) Rabbit mAb (Alexa Fluor® 488 Conjugate) #2216

Applications Reactivity Sensitivity MW (kDa) Isotype
F H M (Mk) (Pg) Endogenous 42 Rabbit IgG

Applications Key:  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  Mk=Monkey  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

PU.1 (9G7) Rabbit mAb (Alexa Fluor® 488 Conjugate) detects endogenous levels of total PU.1 protein. This antibody does not cross react with other Ets family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence of human PU.1 protein. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of MCF-7 cells (blue) or THP-1 cells (green) using PU.1 (9G7) Rabbit mAb (Alexa Fluor® 488 Conjugate).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #2258 reacts with human and mouse PU.1 protein. CST expects that PU.1 (9G7) Rabbit mAb (Alexa Fluor® 488 Conjugate) will also recognize PU.1 in these species.

Background

PU.1 is a member of the Ets family of transcription factors and activates target genes through the purine-rich PU-box (1). PU.1 plays a pivotal role in the differentiation of myeloid cells and lymphocytes and is expressed in several hematopoietic cells including B lymphocytes, macrophages, neutrophils, mast cells, early erythroid cells and megakaryocytes (1,2). The concentration of PU.1 is critical for both the determination of hematopoietic cell lineage and the regulation of differentiation versus stem cell proliferation (3,4). In addition, PU.1 activity is influenced by phosphorylation and interactions with other hematopoietic transcription factors. Phosphorylation of PU.1 at Ser146 by CK2 promotes binding to IRF4 and synergistic activation through the immunoglobulin κ 3' enhancer (5). Treatment of pro-B cells with IL-3 leads to phosphorylation of PU.1 at Ser140, resulting in increased PU.1 activity and activation of the anti-apoptotic gene MCL-1 (6). GATA1 binding blocks PU.1 activity during erythroid cell development (7). Overexpression of PU.1 resulting from proviral insertion during Friend virus infection can induce erythroleukemia, while reduced expression has been associated with acute myeloid leukemia (8).

  1. Lloberas, J. et al. (1999) Immunol. Today 20, 184-189.
  2. Klemsz, M.J. et al. (1990) Cell 61, 113-124.
  3. Dahl, R. and Simon, M.C. (2003) Blood Cells Mol. Dis. 31, 229-233.
  4. DeKoter, R.P. and Singh, H. (2000) Science 288, 1439-1441.
  5. Pongubala, J.M. et al. (1993) Science 259, 1622-1625.
  6. Wang, J.M. et al. (2003) Mol. Cell Biol. 23, 1896-1909.
  7. Zhang, P. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 8705-8710.
  8. Moreau-Gachelin, F. et al. (1998) Nature 331, 277-280.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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