Preparation And Storage
Recommended Assay Procedures
Applications include gel shift, western blot analysis (1-2 µg/ml) and immunoprecipitation. The antibody has been shown to recognize in vitro translated PU.1 and PU.1 expressed as a recombinant protein in bacteria. In. In gel shift assays using µB as a probe (µB is a transcriptional element in the immunoglobulin heavy chain enhancer), G148-74 supershifts the complex of probe and PU.1.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
PU.1 is an ets-related transcription factor expressed in B lymphocytes and macrophages. It was initially identified as the oncogene Spi-1 which was found to block erythroblast differentiation. PU.1 functions by binding transcriptional control elements such as µB in the immunoglobulin heavy chain enhancer and π adjacent to µE2. It has been shown to form a complex with NF-EM5, a B cell transcription factor, resulting in the activation of the immunoglobulin κ 3' enhancer. PU.1 also appears to be critically involved in the control of monocyte development by regulating the expression of the macrophage colony-stimulating factor receptor. PU.1 runs as a 40-42 kDa protein on SDS-PAGE. G148-74 recognizes human PU.1, an Ets-related transcription factor. PU.1 is expressed in B lymphocytes and macrophages and like other Ets-related proteins, binds to consensus sites in DNA through an 85 amino acid Ets domain in the carboxyl terminal region of the protein. The antibody was raised against a bacterially expressed GST-PU.1 fusion protein.
Development References (6)
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Klemsz MJ, McKercher SR, Celada A, Van Beveren C, Maki RA. The macrophage and B cell-specific transcription factor PU.1 is related to the ets oncogene. Cell. 1990; 61(1):113-124. (Biology). View Reference
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Moreau-Gachelin F, Ray D, Mattei MG, Tambourin P, Tavitian A. The putative oncogene Spi-1: murine chromosomal localization and transcriptional activation in murine acute erythroleukemias. Oncogene. 1989; 4(12):1449-1456. (Biology). View Reference
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Pongubala JM, Nagulapalli S, Klemsz MJ, McKercher SR, Maki RA, Atchison ML.. PU.1 recruits a second nuclear factor to a site important for immunoglobulin kappa 3' enhancer activity. Mol Cell Biol. 1992; 12(1):368-378. (Biology). View Reference
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Rivera RR, Stuiver MH, Steenbergen R, Murre C. Ets proteins: new factors that regulate immunoglobulin heavy-chain gene expression. Mol Cell Biol. 1993; 13(11):7163-7169. (Biology). View Reference
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Shin MK, Koshland ME. Ets-related protein PU.1 regulates expression of the immunoglobulin J-chain gene through a novel Ets-binding element. Genes Dev. 1993; 7(10):2006-2015. (Biology). View Reference
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Zhang DE, Hetherington CJ, Chen HM, Tenen DG. The macrophage transcription factor PU.1 directs tissue-specific expression of the macrophage colony-stimulating factor receptor. Mol Cell Biol. 1994; 14(1):373-381. (Biology). View Reference
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