Product Pathways - Lymphocyte Signaling
PU.1 (9G7) Rabbit mAb #2258
|2258S||100 µl (10 western blots)||---||In Stock||---|
|2258||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse||Endogenous||42||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Monkey, Pig.
Specificity / Sensitivity
This antibody detects endogenous levels of total PU.1 protein. The 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.
Western blot analysis of extracts from RAW, THP1 and p388D1 cells using PU.1 (9G7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mouse tumor using PU.1 (9G7) Rabbit mAb #2258.
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma using PU.1 (9G7) Rabbit mAb in the presence of control peptide (left) or PU.1 Blocking Peptide #1036 (right).
Flow cytometric analysis of untreated THP-1 cells using PU.1 (9G7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Confocal immunofluorescent analysis of Raw cells using PU.1 (9G7) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 U-937 cells and either 10 μl of PU.1 (9G7) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human CD11b promoter primers, human CD18 intron 1 primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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).
- Lloberas, J. et al. (1999) Immunol. Today 20, 184-189.
- Klemsz, M.J. et al. (1990) Cell 61, 113-124.
- Dahl, R. and Simon, M.C. (2003) Blood Cells Mol. Dis. 31, 229-233.
- DeKoter, R.P. and Singh, H. (2000) Science 288, 1439-1441.
- Pongubala, J.M. et al. (1993) Science 259, 1622-1625.
- Wang, J.M. et al. (2003) Mol. Cell Biol. 23, 1896-1909.
- Zhang, P. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 8705-8710.
- Moreau-Gachelin, F. et al. (1998) Nature 331, 277-280.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
This antibody is developed, validated, and produced by CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and 7,429,487) from Epitomics, Inc.