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PIF3 | Phytochrome interacting factor 3 (goat antibody)

AS16 3954 | Clonality: Polyclonal | Host: Goat | Reactivity: Arabidopsis thaliana

PIF3 | Phytochrome interacting factor 3 (goat antibody) in the group Antibodies for Plant/Algal  / DNA/RNA/Cell Cycle / Transcription regulation at Agrisera AB (Antibodies for research) (AS16 3954)

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Product Information

Immunogen KLH-conjugated peptide, derived from Arabidopsis thaliana PIF3, UniProt: O80536, TAIR: AT1G09530
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 57 | ca. 65 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Cardamine hirsuta
Not reactive in Nicotiana attenuata

Application examples

Application examples Application example

Western blot using polyclonal anti-PIF3 antibodies

Total protein from Arabidopsis thaliana (10-20 µg) of 4-d-old dark grown Col-0, pif single, triple and quadruple mutants were extracted with the buffer described in Qiu et al. 2015 and denatured at 95°C for 5 min. were separated on 8 % Bis-Tris SDS-PAGE, and subsequently blotted to Nitrocellulose membrane using wet tank transfer. Blots were blocked with TBS containing 2% non-fat milk at room temperature for 1h. Primary anti-PIF3 antibodies were used in dilution of 1:1000 to TBS containing 2% milk. The incubation time was 14-20 hours at 4°C. After washing 4x5 min. with TBST at room temperature, the blots were incubated with HRP-conjugated secondary antibodies (1:5000) at room temperature for 1h. The blots were washed 4x10 min with TBST again and the signal was detected using the SuperSignal West Dura Extended Duration Substrate (ThermoFisher Scientific).

Courtesy od Dr. Yongjian Qiu, University of California-Riverside, USA

Additional information

Additional information

Please, do not re-use PIF3 antibody solution after first incubation with your membrane as most of antibody will bind in this step and next result will not be reproducable.

PIF3 protein runs at higher MW than expected, as observed previously (Al-Sady et al. 2006). 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).


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Plant protein extraction buffer

Secondary antibodies

Background

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,ABI1, ABA INSENSITIVE 1, AtABI1.

Product citations

Selected references

Sinclair et al. (2017) Etiolated Seedling Development Requires Repression of Photomorphogenesis by a Small Cell-Wall-Derived Dark Signal. Curr Biol. 2017 Nov 20;27(22):3403-3418.e7. doi: 10.1016/j.cub.2017.09.063.

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