PhyB | Phytochrome B (dicots)

Product no: AS21 4566

AS21 4566  | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana PhyB, C-terminal, UniProt: P14713, TAIR: At2g18790
    Host: Rabbit
    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: 129 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Arabis alpina, Camelina sativa, Capsella rubella, Brassica napus,  Brassica oleracea, Eutrema salsugineum, Raphanus sativus

    Species of your interest not listed? Contact us
    Not reactive in: Gossypium hirsutum, Hordeum vulgare, Poplar sp., Zea mays
  • Application Examples
  • Western blot using anti-PhyB polyclonal antibodies

    Samples:
    Ler - 30 ug of Arabidopsis thaliana Ler dark grown seedling extract
    phyB-5 - 30 ug of Arabidopsis thaliana phyB-5 mutant dark grown seedling extract
    MW markers: PageRuler Prestained Protein Ladder (Thermo Fisher Scientific)

    30 μg/well of total protein extracted from 4-days-old Arabidopsis thaliana seedlings with extraction buffer (65 mM Tris/HCl pH 6,8; 4 M Urea; 10% glycerol; 3% SDS; 0.05% bromphenol blue; 20 mM DTT). Protein extracts were stored overnight at -20°C. Prior to separation on SDS PAGE, extracts were thawed and incubated for 5 min at 65°C followed by centrifugation and were separated on 9% SDS-PAGE and blotted 2 h to PVDF membrane (Immobilon-P pore size 0.45 μm) using wet transfer. Blot was blocked with 5% milk powder in TBS-T for 1 h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed with TBS-T briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated AS09 602) diluted to 1:25 000 in TBS-T for 1h/RT with agitation. The blot was washed as above and developed for 5 min with AgriseraECLSuperBright with Fusion SL (Peqlab) luminescence camera system. Exposure time was 8 minutes at full resolution.

    Courtesy of Dr. Andreas Hiltbrunner, Freiburg University, Germany

  • Additional Information
  • Additional information: PhyB | Phytochrome B (dicots)
  • Background
  • Background: PhyB (Phytochrome B) is a Red/far-red photoreceptor involved in the regulation of de-etiolation. Protein exists in two inter-convertible forms: Pr and Pfr (active). Involved in the light-promotion of seed germination and in the shade avoidance response. Alternative names: Protein LONG HYPOCOTYL 3, Protein OUT OF PHASE 1, OOP1.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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