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Western blot analysis of TFE3. Lysate from WI-38 cells was probed with anti- TFE3 (clone G138-312) at concentrations of 1.0 (lane 1), 0.5 (lane 2), and 0.25 µg/ml (lane 3). TFE3 is identified as a band of 50 kDa.
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Applications include western blot analysis (0.25-1.0 µg/ml). WI-38 cells are suggested as a positive control. Other applications include gel shift (supershifts) and immunoprecipitation, which are not routinely tested by BD Biosciences Pharmingen. The antibody has been used to detect in vitro translated TFE3-L and TFE3-L, expressed as a recombinant protein in bacteria. In gel shift assays, using µE3 as a probe, this antibody supershifts the complex.
TFE3 is a ubiquitously expressed 50 kDa transcription factor containing both basic helix-loop-helix (bHLH) and leucine zipper (ZIP) motifs. It was initially identified as a protein which bound to the µE3 site in the immunoglobulin heavy chain enhancer. In addition, it binds to the MLTF/USF site in the adenovirus major late promoter. Upon binding as a dimer, it induces a minor groove-oriented bend in the DNA, similar to other HLH proteins such as TFEB, USF, myc, and max. Binding results in transcriptional activation and is mediated through the basic region located just N-terminal to the HLH domain. Clone G138-312 recognizes TFE3. The antibody was raised against a bacterially expressed TFE3-L protein consisting of 327 amino acids encoded by nucleotides 361-2469 of the cloned cDNA.
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.