PIF3 | Phytochrome interacting factor 3

product information background PIF3 (Phytochrome interacting factor 3) is a transcription factor which acts positively in the phytochrome signaling pathway. It activates transcription by binding to the G box. Subcellular localization is nucleus. Alternative names:Basic helix-loop-helix protein 8, AtbHLH8, bHLH8, Phytochrome-associated protein 3, Phytochrome-interacting factor 3, Transcription factor EN 100, EN100, bHLH transcription factor bHLH008, PAP3, PHYTOCHROME-ASSOCIATED PROTEIN 3, POC1, PHOTOCURRENT 1. immunogen KLH-conjugated peptide, derived from Arabidopsis thaliana PIF3, UniProt: O80536, TAIR: AT1G09530 antibody format rabbit polyclonal affinity purified serum lyophilized quantity 50 µg for reconstitution add 50 µl, of sterile water. storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes. tested applications western blot (WB) related products AS12 1860 | anti-PIF4 | PHYTOCHROME INTERACTING FACTOR 4 rabbit antibody AS12 2112 | anti-PIF5 | PHYTOCHROME INTERACTING FACTOR 5 rabbit antibody additional information to be added when available

application information
recommended dilution		1:1000 (WB)
expected | apparent MW		57 | ca. 65 kDa
confirmed reactivity		Arabidopsis thaliana
predicted reactivity		Arabidopsis thaliana
not reactive in		no confirmed exceptions from predicted reactivity known in the moment
additional information		PIF3 protein runs at higher MW than expected, as observed previously (Al-Sady et al. 2006).
selected references		to be added when available, antibody released in April 2013.

application example



Materials: 4-days-old Col-0 and pifq Arabidopsis thaliana seedlings grown under dark (0.2 gram fresh weight) Methods: 0.2 gram of seedlings were extracted by the method described in Galvao et al. (2012). 20 µl of 600 µl plant extracts were separated on 8% Bis-Tris SDS-PAGE and blotted 1h to PVDF membrane. Blots were blocked with 5% milk in TBST for 1 h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for overnight at 4 degree with agitation. The antibody solution was decanted and the blot was washed for 15 min with 3 times with TBST at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG HRP from BioRad) diluted to 1:5000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ThermoScientific PICO Chemiluminescent Substrate. Exposure time was 1 min.

Courtesy Dr. Chan Yul Yoo, Duke University, USA,