TOC1-P | TIMING OF CAB EXPRESSION 1 Arabidopsis clock gene, phosphorylated

Product no: AS22 4701

AS22 4701 | Clonality:  Polyclonal |  Host:  Rabbit | Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana TOC1 phosphorylated protein sequence, UniProt: A0A178UC73, TAIR: AT5G61380
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Antigen affinity purified serum, in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl, of sterile or deionized 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: 69.195 | kDa (due to N-terminal or C-terminal processing)
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-phosphorylated TOC1 antibodies

    8 µg/well of total protein extracted freshly from 10-day-old Arabidopsis thaliana seedlings with extraction buffer (100 mM Tris·Cl, pH 7.5/150 mM NaCl/0.5% NP-40/1mM EDTA/3mM DTT/1 mM PMSF/2 mM NaF/2mM Na3VO4/1 µg ml-1 leupeptin/1 µg ml-1 aprotinin/1 µg ml-1 pepstatin/5 µg ml-1 antipain/5 µg ml-1 chymostatin/50 µM MG132/50 µM MG115/50 µM ALLN) and denatured at room temperature 24 °C for 3 min (TOC1 protein is very labile and sensitive to heat). Proteins were separated on 8% SDS-PAGE regular gels and blotted 1.5h to PVDF (pore size of 0.2 µm), using wet transfer. Blot was blocked with 5% milk for 1.5h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in 5% non-fat milk TBS-T ON/4°C. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed 3 times for 10 min in 5% non-fat milk TBS-T at RT with agitation. Blot was incubated in Agrisera goat anti-rabbit IgG (AS09 602) diluted to 1:25 000 in 5% non-fat milk for 1h/RT. The blot was washed 3 times for 10 min in TBS-T at RT with agitation and developed for 5 min with AgriseraSuperBright (AS16 ECL-S-10). The band marked by a lower arrow is an endogenous TOC1. 

    Courtesy of Dr. David E. Somers, Department of Molecular Genetics, The Ohio State University, USA

  • Additional Information
  • Additional information (application): TOC1 is heat sensitive and requires specific extraction buffer and denaturation conditions, described in application example. Using other conditions, may contribute to lack of detection of phosphorylation of TOC1 using this antibody.
  • Background
  • Background: TOC1 /TIMING OF CAB EXPRESSION 1) is involved in a negative regulation of gene expression and cytokinin activated signaling. Influences carbon fixation and biomass through the circadian clock period.
  • 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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