Auxin response factors (ARFs) are transcription factors that bind to auxin response elements in promoters of early auxin response genes and play central roles in many auxin-mediated processes. As transcription factors, ARFs possess a modular structure and consist of three major domains: an amino-terminal DNA-binding domain (DBD); a middle domain, which is responsible for gene activation or repression; and a carboxy-terminal PB1(Phox and Bem1) domain, contained within a region that was previously called domain III/I. ARFs function as transcription activators or repressors, depending on the middle region of the ARF protein; Q-rich middle regions activate transcription, whereas P/S/T-rich middle regions confer repressor activity.
Background References
1. Guan X et al. Temporal and Spatial Distribution of Auxin Response Factor Genes During Tomato Flower Abscission. Journal of Plant Growth Regulation 33(2):317-327 (2014).
2. John WC. Auxin Response Factors. Plant, Cell and Environment 39:1014-1028 (2016).
Western blot analysis of Auxin response factor on tea leafbud lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:1,000 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded leafbud using anti-Auxin response factor antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1901-06) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. Counter stained with hematoxylin and mounted with DPX.
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