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Purified anti-RPS6 Phospho (Ser235/Ser236) Antibody

Pricing & Availability
Clone
A17020B (See other available formats)
Regulatory Status
RUO
Other Names
S6, 40S ribosomal protein S6
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
A17020B_Pure_RPS6-Phospho_Antibody_1_071318
Total cell lysates (15 µg protein) from serum-starved Jurkat cells treated without (-) or with (+) 160 nM PMA for 15 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with the 1.0 µg/mL (1:500 dilution) of purified anti-RPS6 Phospho (Ser235/Ser236) antibody (Clone A17020B). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal protein loading was confirmed using a pan RPS6 antibody. Lane M: MW ladder.
  • A17020B_Pure_RPS6-Phospho_Antibody_1_071318
    Total cell lysates (15 µg protein) from serum-starved Jurkat cells treated without (-) or with (+) 160 nM PMA for 15 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with the 1.0 µg/mL (1:500 dilution) of purified anti-RPS6 Phospho (Ser235/Ser236) antibody (Clone A17020B). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal protein loading was confirmed using a pan RPS6 antibody. Lane M: MW ladder.
  • A17020B_Pure_RPS6-Phospho_Antibody_2_071318
    Total cell lysates (15 µg protein) from serum-starved NIH/3T3 cells treated without (-) or with 100 ng/mL PDGF-BB (Cat. No. 558804) for 20 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with the 1.0 µg/mL (1:500 dilution) of purified anti-RPS6 Phospho (Ser235/Ser236) antibody (Clone A17020B). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse-IgG (Cat. No. 405301) at a 1:3000 dilution. Equal protein loading was confirmed using a pan RPS6 antibody. Lane M: MW ladder.
  • A17020B_Pure_RPS6-Phospho_Antibody_3_071318
    Serum starved NIH/3T3 cells were untreated (panel A) or stimulated with 100 ng/mL PDGF-BB for 20 minutes. Cell were then fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 1:100 (5 µg/mL) of purified anti-RPS6 Phospho (Ser235/Ser236) antibody overnight at 4°C, after which proteins were visualized with Alexa Fluor® 594 goat anti-mouse IgG (Cat. No. 405308) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • A17020B_Pure_RPS6-Phospho_Antibody_4_071318
    Human peripheral blood lymphocytes were stimulated with (filled histogram) or without (open histogram) Cell Activation cocktail without Brefeldin A (Cat. No. 423302) for 15 minutes, fixed with Fixation Buffer, permeabilied with True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with purified anti-RPS6 Phosopho (Ser235/Ser236) (clone A17020B), followed by anti-mouse IgG PE.
  • A17020B_Pure_RPS6-Phospho_Antibody_5_092222_updated
    IHC staining of purified anti-RPS6 (Ser235/Ser236) (clone A17020B) on formalin-fixed paraffin-embedded human thymus. Following antigen retrieval using 1X Tris-EDTA, pH 9.0, the tissue was treated with (+) or without (-) Lambda Protein Phosphatase overnight, blocked, and incubated with 10.0 µg/mL purified anti-RPS6 (Ser235/Ser236) (Clone A17020B) (panel A) or 5.0 µg/mL of the anti-RPS6 (Cat. 691802) (panel B) overnight at 4°C. This was followed by incubation with Alexa Fluor® 647 goat anti-mouse IgG (Cat. No. 405322) at 2.5 µg/mL. Nuclei were counterstained with DAPI (blue) (Cat. No. 422801) and the slide was mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective. Scale: 50 µm
  • A17020B_Pure_RPS6-Phospho_Antibody_6_072722
    IHC staining of purified anti-RPS6 (Ser235/Ser236) (clone A17020B) on formalin-fixed paraffin-embedded human thymus without DAPI. The image was captured with a 40X objective. Scale: 50 µm.
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608601 25 µg £77
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608602 100 µg £182
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Description

Ribosomal protein S6 (RPS6) is a key component of the small 40S ribosomal subunit and is the major substrate of protein kinases in eukaryotic ribosomes. In response to various cellular stimuli such as mitogenic stimulation, insulin, and increased nutrient availability, upstream kinases such as RSK and p70 kinases phosphorylate RPS6 at multiple serine sites. These modifications facilitate the recruitment of the  7-methylguanasine cap complex, thereby promoting the assembly of the translational pre-initiation complex and increased cellular protein synthesis capacity. RPS6 has been shown to be hyperphosphorylated in certain cancers, and phosphorylation is a critical determinant of pancreatic β–cell size and systemic glucose homeostasis function in diabeteic mouse models.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide from human RPS6 phosphorylated at Serines 235 and 236
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, ICFC, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.1 - 1.0 µg per ml. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/ml is recommended. For intracellular flow cytometry using our True-Phos™ Perm Buffer in Cell Suspension, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µl volume. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration of 10.0 µg/mL is suggested.It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Due to complete conservation of the immunizing sequence between humans, mouse and rat, this clone is is predicted to react with rat RPS6 phosphorylated at serines 235 and 236.

Product Citations
  1. Moore AR, et al. 2022. Cell Rep. 41:111651. PubMed
RRID
AB_2749899 (BioLegend Cat. No. 608601)
AB_2749899 (BioLegend Cat. No. 608602)

Antigen Details

Structure
RPS6 is a 249 amino acid protein with a predicted molecular weight of 28.6 kD.
Distribution

Cytosol, nucleus, ER, ubiquitous expression.

Function
Phosphorylation events at a cluster of serine residues (Ser 235, 236, 240, and 244) at the carboxy-terminus of RPS6 results in increased global translational efficiency. These sites are dephosphorylated during growth arrest.
Interaction
RPS6AK1 and RPS6AK3; other subunits of 40S ribosomal complex.
Biology Area
Cell Biology, Cell Proliferation and Viability, Protein Synthesis, Signal Transduction
Molecular Family
Nuclear Markers, Phospho-Proteins
Antigen References
  1. Jefferies HB, et al. 1997. EMBO J. 16:3693.
  2. Ruvinsky I, et al. 2005. Genes. Dev.19:2199.
  3. Chumacher AM, et al. 2006. Biochemistry. 45:13614
  4. Roux PP, et al. 2007. J. Biol. Chem. 282:14056.
  5. Stevens C, et al. 2009. J. Biol. Chem. 284:334.
  6. Schlafli P, et al. 2011. FEBS J. 278:1757.
Gene ID
6194 View all products for this Gene ID
UniProt
View information about RPS6 Phospho Ser235/Ser236 on UniProt.org
Go To Top Version: 2    Revision Date: 07/27/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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