PIF4 | Phytochrome interacting factor 4 (rabbit)

AS16 3157 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen

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

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4

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 Chromatin immunoprecipitation (ChIP), Western blot (WB)

Recommended dilution 1: 1000 (WB)

Expected | apparent MW 48.3 | 60 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Not reactive in

No confirmed exceptions from predicted reactivity known in the moment

Application examples

Application example

5-20 µg of Arabidopsis thaliana nuclear proteins were extracted according to Gendrel et al. 2005. Samples were denatured with SDS page loading buffer at 90°C for 5 min, separated on 12% SDS-PAGE and then blotted 1h to PVDF using wet transfer. Blot was blocked with 5% milk 1h at RT with agitation and then It was incubated with the anti-PIF4 antibody at a dilution of 1:1000 in TBS-T ON at 4°C in agitation. The antibody solution was decanted and the blot was rinsed briefly with TBS-T, then washed 3 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in TBS-T for 1h/RT with agitation (Agrisera AS09 602). The blot was washed as above and developed with chemiluminescent ragent. Exposure time was 30 min.

Courtesy of Dr. Lucia Ferrero, Universidad Nacional del Litoral, Argentina

Additional information

PIF proteins are very unstable, therefore special precautions should be taken during extraction and whole procedure should be performed in the dark and with as little light as possible (green light only).

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

This product can be sold containing ProClin if requested.

Related products

Plant protein extraction buffer

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 Gras et al. (2020). Arabidopsis Thaliana SURFEIT1-like Genes Link Mitochondrial Function to Early Plant Development and Hormonal Growth Responses. Plant J . 2020 Apr 5. doi: 10.1111/tpj.14762.
Ferrero et al. (2019). Class I TCP transcription factors target the gibberellin biosynthesis gene GA20ox1 and the growth promoting genes HBI1 and PRE6 during thermomorphogenic growth in Arabidopsis. Plant Cell Physiol. 2019 Jul 11. pii: pcz137. doi: 10.1093/pcp/pcz137.
Hwang et al. (2019). Trehalose-6-phosphate signaling regulates thermoresponsive hypocotyl growth in Arabidopsis thaliana. EMBO Rep. 2019 Aug 8:e47828. doi: 10.15252/embr.201947828.