Technical Data
P9190
PU.1 (Ets Transcription Factor)
Description:
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 Casein Kinase II (CKII) promotes binding to IRF4 and synergistic activation through the immunoglobulin k 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).

Applications:
Suitable for use in ELISA, Immunofluorescence, Flow Cytometry, Western Blot, Immunoprecipitation, Immunohistochemistry and Immunocytochemistry. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:1000
Immunoprecipitation: 1:100
Chromatin Immunoprecipitation: 1:25
Immunohistochemistry (Paraffin): 1:200. Antigen retrieval: heat in citrate buffer, pH 6.0.
Immunofluorescence (IF-IC): 1:100
Flow Cytometry: 1:50
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceHumanRabbit
Concentration:
Not determined
Immunogen:
Synthetic peptide corresponding to amino acids at the amino-terminus of human PU.1 (KLH).
Purity:
Purified by Protein A and peptide affinity chromatography.
Form
Supplied in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity:
Recognizes endogenous levels of total human PU.1 protein at ~42kD. Does not cross react with other Ets family members. Species Crossreactivity: mouse. Species Sequence Homology: monkey and porcine.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Lloberas, J. et al. (1999) Immunol. Today 20, 184189. 2. Klemsz, M.J. et al. (1990) Cell 61, 113124. 3. Dahl, R. and Simon, M.C. (2003) Blood Cells Mol. Dis. 31, 229233. 4. DeKoter, R.P. and Singh, H. (2000) Science 288, 14391441. 5. Pongubala, J.M. et al. (1993) Science 259, 16221625. 6. Wang, J.M. et al. (2003) Mol. Cell. Biol. 23, 18961909. 7. Zhang, P. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 87058710. 8. Moreau-Gachelin, F. et al. (1998) Nature 331, 277280.