RPS6A-P240 | Phosphorylated (Ser240) 40S ribosomal protein S6-1
AS19 4302 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
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 ĩg
Reconstitution
For reconstitution add 50 ĩl, of sterile water
Storage
Store lyophilized/reconstituted at -20°C; 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
Species of your interest not listed? Contact us
Application examples
Application examples
Lanes:
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.
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
Additional information
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
To be added when available, antibody released in February 2021.
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