PU.1 (9G7) Rabbit mAb (Alexa F

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PU.1 (9G7) Rabbit mAb (Alexa Fluor^® 647 Conjugate) #2240

Applications	Reactivity	Sensitivity	Isotype
F	H M	Endogenous	Rabbit IgG

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

Protocols

2240:

Flow

Specificity / Sensitivity

PU.1 (9G7) Rabbit mAb (Alexa Fluor^® 647 Conjugate) 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. The antibody was conjugated to Alexa Fluor^® 647 under optimal conditions with an F/P ratio of 2-6. The Alexa Fluor^® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor^® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor^® 647 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^® 647 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

• 2216 PU.1 (9G7) Rabbit mAb (Alexa Fluor^® 488 Conjugate)
• 2258 PU.1 (9G7) Rabbit mAb
• 2266 PU.1 Antibody
• 2985 Rabbit (DA1E) mAb IgG XP^® Isotype Control (Alexa Fluor^® 647 Conjugate)
• 4964 IRF-4 Antibody

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

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