PIF4 | Phytochrome interacting factor 4 (goat antibody)

AS16 3955 | Clonality: Polyclonal | Host: Goat | Reactivity: Arabidopsis thaliana

Product Information

Immunogen KLH-conjugated peptide derived from Arabidopsis thaliana PIF4, UniProt:Q8W2F3, TAIR:AT2G43010

Host Goat

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

Format Lyophilized

Quantity 50 µg

Reconstitution 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)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW 48.3 | 60 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Brassica napus, Brassica rapa, Camelina sativa, Eutrema salsugineum
Species of your interest not listed? Contact us

Not reactive in Solanum lycopersicum

Application examples

Application examples Application example

30 ul of total protein from Arabidopsis thaliana grown in 12 hour light/ 12 hour dark growth conditions and 6-day-old seedling samples taken at the end of the day (ZT12) were extracted with 100 mM MOPS, pH 7.6, 100 mM NaCl, 10% glycerol, 40 mM 2-mercaptoethanol, 5% SDS, 1X protease inhibitor cocktail from Roche, 2 mM PMSF. 80 µl buffer were added to about 100 µl grinded powder, then immediately heated at 70ºC for 10 minutes and separated on 8 % Bis-Tris SDS-PAGE and blotted 1h to PVDF using semi-dry or tank transfer. Blots were blocked with 5 % non-fat milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for over night at 4ªC with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-goat IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 minutes with chemiluminescent detection reagent. Exposure time was 1 minute.

Courtesy of Dr. Bo Zhang, Umeå Plant Science Centre, Sweden

Additional information

PIF proteins are not that stable, therefore special precautions should be taken during extraction and whole procedure should be performed in as little light as possible (light green light).

Extraction of PIF proteins is described in Shen et al. (2007).

Material used need to be up to 8 days old as detection in older rosette leaf may fail.

Related products

Plant protein extraction buffer

Secondary antibodies

Background

Background PIF4 (PHYTOCHROME INTERACTING FACTOR 4) is a transcription factor involved in the phytochrome B signaling pathway. Interacts with APRR1/TOC1 and PIF3 and binds to EGL2 and RGA. Alternative names: AtPIF4, Basic helix-loop-helix protein 9, AtbHLH9, bHLH 9, SRL2, Short under red-light 2, EN102, Transcription factor EN 102, bHLH transcription factor bHLH009.

Product citations

Selected references Lee, Paik & Huq. (2020). SPAs promote thermomorphogenesis by regulating the phyB-PIF4 module in Arabidopsis. Development. 2020 Oct 8;147(19):dev189233. doi: 10.1242/dev.189233. PMID: 32994167; PMCID: PMC7561471.
Sun et al. (2019). SHB1 and CCA1 interaction desensitizes light responses and enhances thermomorphogenesis. Nat Commun. 2019 Jul 15;10(1):3110. doi: 10.1038/s41467-019-11071-6.