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RPS6A-P237 | Phosphorylated (Ser237) 40S ribosomal protein S6-1

AS19 4291   | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Arabidopsis thaliana

RPS6A-P237 | Phosphorylated (Ser237) 40S ribosomal protein S6-1 in the group Antibodies for Plant/Algal  / DNA/RNA/Cell Cycle / Translation at Agrisera AB (Antibodies for research) (AS19 4291)

DATA SHEET IN PDF

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Product Information

Immunogen KLH-conjugated peptide derived from Arabidopsis thaliana RPS6A with phosphorylated Ser237, UniProt: O48549, TAIR: At4g31700
Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum, in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, of sterile 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 28.3 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
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples

Western blot using anti-RPS6A-P237 antibodies

Left panel is developed with anti-RPS6-Ser237,  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


Western blot using anti-RPS6A-P237 antibodies


20 µg/well of total protein freshly extracted freshly from 16 d old Arabidopsis thaliana seedlings total leaf of wildtype (1) and mutant (2) with 2x SDS loading dye exact buffer and denatured at 80 °C for 5 min.were separated on 12% SDS-PAGE  and blotted 10 min to PVDF (pore size of 0.2 um), using using Turbo transfer (BioRad). Blot was blocked with 5% milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly once, then washed 5 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:15 000 in  for 1h/RT with agitation. The blot was washed as above and reaction was visualized with chemiluminescence, following manufacture's instructions. Exposure time was 5 seconds.

Courtesy of Dr. Arsheed Sheikh, King Abdullah University of Science and Technology (KAUST), Saudi Arabia

Additional information

Related products

Related products AS19 4292 | Anti-RPS6A | 40S ribosomal protein S6-1, rabbit antibodies

collection of antibodies involved in translation

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 November 2020.

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