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Purified anti-RPS6 Phospho (Ser244) Antibody

Pricing & Availability
Clone
A18024A (See other available formats)
Regulatory Status
RUO
Other Names
Ribosomal Protein S6, 40S Ribosomal Protein S6, Phosphoprotein NP33, Small Ribosomal Subunit Protein ES6
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
A18024A_PURE_RPS6-Phospho_Antibody_1_030320
Total cell lysates (15 µg protein) from serum-starved SR cells treated without (-) or with (+) 160 nM PMA for 30 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2500 dilution) of purified anti-RPS6 Phospho (Ser244) antibody (clone A18024A). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Equal protein loading was confirmed using a pan RPS6 antibody and Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) used at a 1:25000 dilution (lower). Lane M: molecular weight ladder.
  • A18024A_PURE_RPS6-Phospho_Antibody_1_030320
    Total cell lysates (15 µg protein) from serum-starved SR cells treated without (-) or with (+) 160 nM PMA for 30 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2500 dilution) of purified anti-RPS6 Phospho (Ser244) antibody (clone A18024A). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Equal protein loading was confirmed using a pan RPS6 antibody and Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) used at a 1:25000 dilution (lower). Lane M: molecular weight ladder.
  • A18024A_PURE_RPS6-Phospho_Antibody_2_030320
    Total cell lysates (15 µg protein) from serum-starved NIH/3T3 cells treated without (-) or with (+) 20% FBS for 30 minutes were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 0.25 µg/mL (1:2000 dilution) of purified anti-RPS6 Phospho (Ser244) antibody (clone A18024A). Proteins were visualized by chemiluminescence detection using HRP goat anti-mouse IgG antibody (Cat. No. 405306) at a 1:3000 dilution. Equal protein loading was confirmed using a purified anti-RPS6 antibody (Cat. No. 691802) and Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) used at a 1:25000 dilution (lower). Lane M: molecular weight ladder.
  • A18024A_PURE_RPS6-Phospho_Antibody_3_030320
    Serum starved NIH/3T3 cells were untreated (panel A) or treated with 20% FBS for 30 minutes. Cells were then fixed with 4% paraformaldehyde for 10 minutes, permeabilized with methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 2 µg/mL of purified anti-RPS6 Phospho (Ser244) antibody (clone A18024A) overnight at 4°C, after which proteins were visualized with Alexa Fluor® 594 goat anti-mouse IgG antibody (Cat. No. 405326) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • A18024A_PURE_RPS6-Phospho_Antibody_4_030320
    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, permeabilized with True-Phos™ Perm Buffer (Cat. No. 425401), and intracellularly stained with purified anti-RPS6 Phospho (Ser244) (clone A18024A), followed by anti-mouse IgG PE.
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935701 25 µg 112 CHF
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935702 100 µg 265 CHF
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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 diabetic mouse models.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide corresponding to human RPS6 phosphorylated at serine 244
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 - 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/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, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

BioLegend offers two clones against this target, A18024A and A18024B:

For western blotting, A18024B displayed a higher affinity for RPS6 Phospho (Ser244) compared to A18024A. Both clones exhibit mouse reactivity.

For ICC, A18024B displayed a moderately higher affinity for RPS6 Phospho (Ser244) compared to A18024A. Both clones are mouse reactive for this application, and both clones are compatible with Triton X-100 and methanol permeabilization steps.

For ICFC, A18024A works in all three ICFC buffers (Cat# 425401, 421002, 424401). For ICFC, A18024A weakly stains mouse RPS6 Phospho (Ser244). A18024B is not recommended for ICFC due to high background staining.

Both clones are predicted to react with rat RPS6 when phosphorylated at serine 244 due to complete sequence homology between the immunizing sequence and the rat RPS6 ortholog.

RRID
AB_2832900 (BioLegend Cat. No. 935701)
AB_2832900 (BioLegend Cat. No. 935702)

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
Regulation of protein translation
Biology Area
Cell Biology, Cell Proliferation and Viability, Protein Synthesis, Signal Transduction
Molecular Family
Organelle Markers, Phospho-Proteins
Antigen References
  1. Jefferies HB, et al. 1997. EMBO J. 16:3693.
  2. Ruvinsky I, et al. 2005. Genes Dev. 19:2199-211.
  3. Schumacher 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 on UniProt.org
Go To Top Version: 2    Revision Date: 01.05.2021

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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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