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



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 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 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

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.
immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana RPS6A with phosphorylated Ser237, UniProt: O48549, TAIR: At4g31700
Reconstitution: For reconstitution add 50 ĩl, of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 ĩg
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)
calculated | apparent molecular mass [kDa]: 28,3 kDa
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
Picture (footer):

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

background: RPS6A (40S ribosomal protein S6-1) is the major substrate of protein kinases in eukaryote ribosomes. Essential during gametogenesis.
All references: To be added when available, antibody released in November 2020.

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