S6K1/2 | Ribosomal S6 kinase 1/2

AS12 1855 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen

KLH-conjugated peptide, derived from Arabidopsis thaliana S6K1: UniProt: P42818, TAIR: AT3G08730 and S6K2: UniProt: Q39030, TAIR: AT3G08720. Due to high amino acid homology, chosen peptide is conserved in both proteins: S6K1 and S6K2.

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum in PBS, pH 7.4 and 0.02% sodium azide.

Format Lyophilized

Quantity 50 µg

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 : 750-1 : 1000 (WB)

Expected | apparent MW

52.6 kDa (S6K1) and 53 kDa (S6K2)

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Thelungiella halophila
Species of your interest not listed? Contact us

Not reactive in Setaria viridis

Application examples

Application examples application example

western blot using anti-S6K/2 antibodies

20 µg of total protein from flowers and leaves of transgenic Arabidopsis thaliana lines were analysed (expressing the genomic copy of S6K1 tagged with FLAG epitop under the control of its own promoter) extracted with homogenization buffer were separated on 10% SDS-PAGE and blotted 2h to PVDF. Blots were blocked with 5% milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:750 for overnight at 4C with agitation. The antibody solution was decanted and the blot was washed three times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated from Agrisera, AS09 602) diluted to 1:10 000 in 2.5% milk in TBST 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 few minutes.

Courtesy of Dr. Rossana Henriques, CRAG, Spain

western blot using anti-S6K polyclonal antibodies

western blot using anti-S6K polyclonal antibodies

20 µg of total protein from Arabidopsis thaliana total wilde type (Col-0), deletion mutant (s6k1-1), overexpression mutant (S6K1g-CFP) extracted with homogenization buffer were separated on 10% SDS-PAGE and blotted 2h to PVDF. Blots were blocked with 5% milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for overnight at 4C with agitation. The antibody solution was decanted and the blot was washed three times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated from Agrisera, AS09 602) diluted to 1:10 000 in 2.5% milk in TBST 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 few minutes.

Courtesy of Dr. Rossana Henriques, CRAG, Spain

Additional information

Additional information So far in applied conditions and extracts endogenous S6K1/2 protein is detectable as a very weak band.

Keeping the samples at 4°C all times is of crucial importance. Laucs buffer was used in example below since this is the one which was used routinely by this test laboratory for the work with kinases. Some unspecific bands can be seen, depending upon western blot protocol which is used. Therefore please consider to use a negative control together with your samples.

Related products

AS12 2608 | Anti-TOR | Target of rapamycin, rabbit antibodies
AS13 2664 | Anti-S6K1-2 | Ribosomal-protein S6 kinase homolog 1,2 - phosphorylated, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

Background

Ribosomal S6 kinase 1/2 (S6K1/2) involved in TOR signaling pathway, in osmotic stress response. Activated by PDK1 and repressed during osmotic stress. Expressed in all tissues, especially during high metabolic activity in growing buds, root tips, leaf margins and germinating seeds. Alternative names: AtPK1/AtPK6 S6K1), AtPK2/AtPK19 (S6K2).

Product citations

Selected references González-López et al. (2021). Growth promotion in Arabidopsis thaliana by bacterial cyclodipeptides involves the TOR/S6K pathway activation. Journal of Plant Physiology. Volume 257, 2021, 153343,ISSN 0176-1617, https://doi.org/10.1016/j.jplph.2020.153343.
Kazibwe et al. (2020). TOR mediates the autophagy response to altered nucleotide homeostasis in a ribonuclease mutant. J Exp Bot. 2020 Sep 9;eraa410.doi: 10.1093/jxb/eraa410.
Dong et al. (2019). The Arabidopsis THADA homologue modulates TOR activity and cold acclimation. Plant Biol (Stuttg). 2019 Jan;21 Suppl 1:77-83. doi: 10.1111/plb.12893.
Inaba and Nagy (2018). Tombusvirus RNA replication depends on the TOR pathway in yeast and plants. Virology. 2018 Jun;519:207-222. doi: 10.1016/j.virol.2018.04.010.
Pfeiffer et al. (2016). Integration of light and metabolic signals for stem cell activation at the shoot apical meristem. Elife. 2016 Jul 11;5. pii: e17023. doi: 10.7554/eLife.17023.