SGS3 | Protein suppressor of gene silencing 3

Product no: AS15 3099

AS15 3099  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen:
    KLH-conjugated peptide derived from Arabidopsis thaliana SGS3 sequence, Uniprot: Q9LDX1, TAIR: AT5G23570
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Immunogen affinity purified serum in PBS pH 7.4.
    Format: Lyophilized
    Quantity: 50 g
    Reconstitution: For reconstitution add 50 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1: 1000 (WB)
    Expected | apparent MW: 72 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Arabidopsis lyrata, Brassica napus, Brassica oleracea, Brassica rapa, Camelina sativa, Raphanus sativus
    Species of your interest not listed? Contact us
    Not reactive in:

    Zea mays

  • Application Examples
  •  Application example

    Western blot using anti-SGS3 antibody
    
    250 µg of grounded tissue from 2 weeks old Arabidopsis thaliana flowers from wt Col-0 and sgs3 mutant was incubated with 1 mL lysis buffer (50 mM HCl pH=7.5, 150 mM NaCl, 10% glycerol, 5mM MgCl2, 0,1% Nonidet P40, Roche protease inhibitors, 1 mM TCEP) and denatured with Laemmli buffer (2% SDS, 10% glycerol, 0.125 M Tris-HCl pH=6.8, 1% DTT) at 85°C for 7 min. The samples were separated on 12% SDS-PAGE and blotted for 60 min to nitrocelulose membrane using wet transfer. Blots were blocked with 5% milk powder  in PBST for 180 min in a cold room with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for O/N in a cold room with agitation. The antibody solution was decanted and the blot was rinsed briefly once, then washed 5 times in TBST/PBST buffer at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera) diluted to 1:10 000 in for 60 min at RT with agitation. The blot was washed as above and developed for 1 min with (ECL DIY mix (100 mM Tris pH8.8, 2 mM 4IBPA (SIGMA), 1.25 mM Lumminol (SIGMA)) using X-ray film. Exposure time was 60 seconds.

    Courtesy of Maria Louisa Vigh, Department of Computational and RNA Biology, University of Copenhagen, Denmark

     

  • Background
  • Background:

    Protein supressor of gene silencing 3 (SGS3) is a protein which is necessary for post-transcriptional gene silencing and virus resistance.

  • Product Citations
  • Selected references: Sun et al. (2021) The epigenetic factor FVE orchestrates cytoplasmic SGS3-DRB4-DCL4 activities to promote transgene silencing in Arabidopsis. Sci Adv. 2021 Aug 4;7(32):eabf3898. doi: 10.1126/sciadv.abf3898. PMID: 34348894; PMCID: PMC8336953.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

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This product can be purchased in 3 different volumes:

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