Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Xenograft

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all Ribosomal Protein S6/RPS6 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing human, mouse and rat Ribosomal Protein S6 S235/S236 sites

Specificity

Detects human, mouse, and rat ribosomal protein S6 when dually phosphorylated at S235 and S236.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images

Ribosomal Protein S6 antibody in Human Colon Cancer Tissue by Immunohistochemistry (IHC-P).

Ribosomal Protein S6 in Human Colon Cancer Tissue.

Ribosomal Protein S6 was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using 5 µg/mL Rabbit Anti-Human/Mouse/Rat Phospho-Ribosomal Protein S6 (S235/S236) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3918) overnight at 4 °C. Tissue was stained with the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific labeling was localized to vascular endothelial cell within glomeruli. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human and Mouse Phospho-Ribosomal Protein S6 (S235/S236) antibody by Western Blot.

Detection of Human and Mouse Phospho-Ribosomal Protein S6 (S235/S236) by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-1 (Catalog # 291-G1) for 20 minutes and NIH-3T3 mouse embryonic fibroblast cell line untreated or treated with 10 ng/mL Human PDGF (Catalog # 120-HD) for 20 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Ribosomal Protein S6 (S235/S236) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3918), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Ribosomal Protein S6 (S235/S236) at approximately 32 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-Ribosomal Protein S6/RPS6 (S235/S236) antibody by Simple WesternTM.

Detection of Human Phospho-Ribosomal Protein S6/RPS6 (S235/S236) by Simple WesternTM.

Simple Western lane view shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human IGF-I (Catalog # 291-G1) for 20 minutes, loaded at 0.5 mg/mL. A specific band was detected for Phospho-Ribosomal Protein S6/RPS6 (S235/S236) at approximately 40 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3918). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human Ribosomal Protein S6/RPS6 by Western Blot

Detection of Human Ribosomal Protein S6/RPS6 by Western Blot

ALM inhibits HIF-1 alpha translation by down-regulating mTOR pathway. (A) Hep3B and PC3 cells were exposed to vehicle or the hypoxia mimics dimethyloxalylglycine (DMOG), cobalt chloride (CoCl2), desferrioxamine (DFX) or MG132 in the presence of ALM at indicated concentrations or vehicle for 24 h and whole cell lysates were subjected to Western blot. (B) Hep3B cells were pretreated for 4 h with vehicle (Vhc), 25 nM rapamycin (Rapa), 20 μg/mL cycloheximide (Chx), or 200 nM ALM, [35S] methionine/cysteine was added for 1 h followed by cell lysis, HIF-1 alpha immunoprecipitation (IP), SDS/PAGE, and autoradiography. (C–E) PC3 cells were cultured at 20% or 1% O2 for 24 h in the presence of ALM (at the indicated concentrations) and whole cell lysates were subjected to for: (C) p-P70, p-Akt, p-mTOR (2448/2481), mTOR, (D) p-4E-BP1, (E) p-RPS6, HIF-1 alpha, or beta -actin. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31083403), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon cancer tissue

Simple Western

5 µg/mL
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1)

Western Blot

0.5 µg/mL
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑1 (Catalog # 291-G1) and NIH‑3T3 mouse embryonic fibroblast cell line treated with Human PDGF (Catalog # 120-HD)

Reviewed Applications

Read 2 reviews rated 4.5 using AF3918 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Ribosomal Protein S6/RPS6

40S ribosomal protein S6 is the major substrate of protein kinases, particularly p70 S6 kinase, in eukaryotic ribosomes. S6 phosphorylation at S235, S236, S240, and S244 upregulates the translation of mRNAs containing an oligopyrimidine tract at their transcriptional start sites. This phosphorylation is stimulated by growth factors, tumor promoting agents, and other mitogens.

Alternate Names

RPS6

Entrez Gene IDs

6194 (Human); 20104 (Mouse); 29304 (Rat)

Gene Symbol

RPS6

Additional Ribosomal Protein S6/RPS6 Products

Product Documents

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices

For research use only

Related Research Areas

Interferons
Translation Regulation

Citations for Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antibody

Customer Reviews (2)

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2 Customer Ratings
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  • Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Bone marrow cells
    Species: Mouse
    Verified Customer | Posted 11/30/2022
    Mouse bone marrow stromal cells were lysed with NP-40 buffer for protein extraction. 10 ug of proteins were loaded in a 10% Acrylamide gel.
    Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antibody AF3918
  • Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antibody
    Name: Francesca Pacifici
    Application: Western Blot
    Sample Tested: Skeletal muscle tissue
    Species: Mouse
    Verified Customer | Posted 11/20/2017
    Western Blot: Phospho S6/RPS6 [AF3918] - Total protein from mouse skeletal muscle tissue, separated on a 4-12% gel by SDS-PAGE, transferred to nitrocellulose membrane and blocked in 5% non-fat milk for 1h at room temperature. The membrane was probed with anti-phospho S6/RPS6 1:400 in non-fat milk.
    Human/Mouse/Rat Phospho-Ribosomal Protein S6/RPS6 (S235/S236) Antibody AF3918

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