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HY5 | Protein long hypocotyl 5

272 €
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AS12 1867 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana

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AS12 1867

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product information
Background HY5 (Protein long hypocotyl 5) is a light-regulated transcription factor that promotes photomorphogenesis in light. Involved in the blue light specific pathway. Heterodimer. Expressed in root, hypocotyl, cotyledon, leaf, stem and floral organs. Localized in nucleus. Alternative names: Protein LONG HYPOCOTYL 5, bZIP transcription factor 56, AtbZIP56.
Immunogen KLH-conjugated peptide, derived from Arabidopsis thaliana HY5 protein sequence, UniProt:O24646 , TAIR: AT5G11260. Chosen peptide is not conserved in HYH protein sequence.
Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized 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 Western blot (WB)
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Additional information
application information
Recommended dilution 1 : 1000 (WB)
Expected | apparent MW

18.5 kDa

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Brassica pekinensis
Not reactive in

Oryza sativa, Pisum sativum, Populus sp. , Solanum lycopersicum

Additional information

This antibody detects well recombinant HY5 in Nicotiana benthamiana (image below) while a signal in endogenous total cell extract of Arabidopsis thaliana has not been optimised in applied conditions (as below).

Using nuclear fraction for detection of endogenous protein is recommened, however it has not been optimized yet.

Selected references Lee et al. (2017). The F-box protein FKF1 inhibits dimerization of COP1 in the control of photoperiodic flowering. Nat Commun. 2017 Dec 22;8(1):2259. doi: 10.1038/s41467-017-02476-2.

application information




western blot using anti-HY5 antibodies

20 ug of total protein from control (1), 35S::YFP-HY5-HA (2, red arrow), 35S::YFP-HY5-HA + 35S::CFP-X protein (green arrow), were separated on 12 % SDS-PAGE using tank transfer and blotted 1 h to PVDF (Biorad). Blots were blocked with 5 % skim milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HY5 antibody (second panel from the left) at a dilution of 1: 1000 for 1h at RT with agitation. 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 PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG, HRP conjugated from Agrisera, AS09 602), diluted to 1: 10 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min. with ECL according to the manufacturer's instructions. Exposure time was 60 seconds.

Courtesy of Dr. Seok Keun Cho, University of Copenhagen, Danmark

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