Phospho-S6 Ribosomal protein (Ser235) Monoclonal antibody

Phospho-S6 Ribosomal protein (Ser235) Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

2A4B6

Cat no : 67898-1-Ig

Synonyms

P RPS6 Ser235, P RPS6 (Ser235), P RPS6, P S6 Ribosomal protein (Ser235), Phospho-RPS6

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Validation Data Gallery

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  WB analysis using 67898-1-Ig

  Non-treated and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67898-1-Ig (Phospho-S6 Ribosomal protein (Ser235) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with beta tubulin antibody as loading control.

• 

  WB analysis using 67898-1-Ig

  Non-treated and Calyculin A treated cells were subjected to SDS PAGE followed by western blot with 67898-1-Ig (Phospho-S6 Ribosomal protein (Ser235) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with beta tubulin antibody as loading control.

• 

  WB analysis using 67898-1-Ig

  Non-treated and IGF-1 treated MCF-7 cells were subjected to SDS PAGE followed by western blot with 67898-1-Ig (Phospho-S6 Ribosomal protein (Ser235) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with beta tubulin antibody as loading control.

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  IF Staining of HeLa using 67898-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed untreated and 100 nM Calyculin A (30 minutes) treated HeLa cells using Phospho-S6 Ribosomal protein (Ser235) antibody (67898-1-Ig, Clone: 2A4B6) at dilution of 1:2000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

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  FC experiment of HeLa using 67898-1-Ig

  1x10^6 untreated HeLa cells (dash lines) and 100 nM Calyculin A (30 minutes) treated HeLa cells (full lines) were intracellularly stained with 0.2 μg Phospho-S6 Ribosomal protein (Ser235) Monoclonal antibody (67898-1-Ig, Clone: 2A4B6, red) and Multi-rAb CoraLite ® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM005). Mouse IgG2b isotype control Mouse McAb (66360-3-Ig, Clone: 11B8C4, blue) was parallel stained as control. Cells were fixed with 4% PFA .

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, Calyculin A treated HeLa cells, Calyculin A treated HEK-293 cells, IGF-1 treated MCF-7 cells, Calyculin A treated NIH/3T3 cells
• Positive IF/ICC detected in: 100 nM Calyculin A (30 minutes) treated HeLa cells
• Positive FC (Intra) detected in: 100 nM Calyculin A (30 minutes) treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:1000-1:4000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below

Product Information

67898-1-Ig targets Phospho-S6 Ribosomal protein (Ser235) in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: ribosomal protein S6
• Calculated Molecular Weight: 29 kDa
• Observed Molecular Weight: 32 kDa
• GenBank Accession Number: BC000524
• Gene Symbol: RPS6
• Gene ID (NCBI): 6194
• RRID: AB_2918654
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Ribosomal protein S6 (RPS6) is one of the components of the 40S ribosomal subunit. RPS6 has been functionally regarded as the stimulator and/or inhibitor of certain types of mRNA translation, as well as the regulator of cellular metabolisms, cells size, survival and proliferation. RPS6 is phosphorylated at multiple sites, comprised between Ser235 and Ser247, by the p70 rpS6 kinase (S6K) 1, which is a major downstream effector of the mammalian target of rapamycin complex 1 (mTORC1). Phosphorylation of RPS6 at the dual site Ser235/236 occurs also independently of mTORC1, via the p90 ribosomal S6 kinases (RSK), which are activated by the extracellular signal-regulated kinases (ERK). Recent studies performed in pancreatic β-cells identified PKA as an additional RPS6 kinase, specifically involved in the phosphorylation of Ser235/236. (PMID: 26490682, PMID: 21814187, PMID: 31112404). 67898-1-Ig specifically recognizes the phosphorylation site of Ser235 or dual site Ser235/236.

Publications

Species	Application	Title
human	WB	Nat Commun; Pharmacological inhibition of Lin28 promotes ketogenesis and restores lipid homeostasis in models of non-alcoholic fatty liver disease; Authors - Evangelia Lekka
human	WB	FASEB J; Pharmacological activation of ERβ by arctigenin maintains the integrity of intestinal epithelial barrier in inflammatory bowel diseases.; Authors - Yu Tao