RPS6A-P240 | Phosphorylated (Ser240) 40S ribosomal protein S6-1

Product no: AS19 4302

AS19 4302   | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Arabidopsis thaliana

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana RPS6A, UniProt: O48549, TAIR: At4g31700 phosphorylated at Ser240
    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: kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana
    Predicted reactivity: Actinidia rufa, Ananas comosus, Beta vulgaris, Cajanus cajan, Capsicum chinense, Citrus clementina, Gossypium australe, Olea europaea subsp. europaea, Oryza sativa, Panicum miliaceum, Populus alba, Sesamum indicum, Senna tora, Solanum lycopersicum, Vigna unguiculata

    Species of your interest not listed? Contact us
  • Application Examples
  • Western blot using anti-RPS6A/P240 antibodies

    Left panel is developed with anti-RPS6-p240,  while right panel anti-RPS6, which serves as a loading control.


    Lanes: 

    1 and 4 - 0.5 µg of Arabidopsis thaliana whole leaf extract from untreated leaf discs

    2 and 5 - 0.5 µg of Arabidopsis thaliana whole leaf extract from leaf discs treated with 10 mM glutamine (TOR activator) for 8 hours

    3 and 6 - 0.5 µg of Arabidopsis thaliana whole leaf extract from leaf discs treated with 2 µM AZD-8055 (TOR inhibitor) for 8 hours


    0.5 µg/well of total protein extracted freshly from mature Arabidopsis thaliana leaves  with 50 mM HEPES pH 7.5, 5mM NaF, 2.5 mM NaPPi, 25 mM B-phosphoglycerol, Roche Complete inhibitor tablet (1x), 2% PVPP, 2 mM PMSF) and denatured with 2x SDS sample buffer (80 mM tris pH 6.8, 2% SDS, 10% glycerol, 100 mM DTT, bromophenol blue) at 99°C for 3 min. were separated on AnyKd (BioRAD) gradient % SDS-PAGE  and blotted 1h to PVDF (pore size of  0.45  um) using semi-dry transfer. Blot was blocked with 2% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in TBS-T + 2% milk powder ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 5 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 + 2 % milk powder  for 1h/RT with agitation. The blot was washed as above and developed for 5  min with Agrisera ECLBright with  GE Amersham Imager 600. Exposure time was ~60.seconds.

    Courtesy of Dr. Brendan O'Leary, University of Western Australia, School of Molecular Sciences, ARC Centre for Plant Energy Biology, Australia

  • Background
  • Background: RPS6A  (40S ribosomal protein S6-1)  is the major substrate of protein kinases in eukaryote ribosomes. Essential during gametogenesis.
  • Product Citations
  • Selected references: 38221900
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts


    Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela

    Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela
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