Human/Mouse/Rat p70 S6 Kinase Antibody

(2 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat p70 S6K and p85 S6K (also known as p70 S6K alpha I), an isoform with 23 extra residues at the N-terminus. Reactivity with  beta isoforms of p70 S6K is unknown.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human p70 S6K
    Accession # M60725
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.2 µg/mL
    See below
  • Simple Western
    2 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Mouse p70 S6 Kinase by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line, HeLa human cervical epithelial carcinoma cell line, and NIH‑3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human/Mouse/Rat p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for p70 S6 Kinase at approximately 70 and 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Intracellular Staining by Flow Cytometry
Detection of p70 S6 Kinase in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Rabbit Anti-Human/Mouse/Rat p70 S6 Kinase Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF8962, filled histogram) or control antibody (Catalog # AB‑105‑C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Immunohistochemistry
p70 S6 Kinase in Human Breast Cancer Tissue. p70 S6 Kinase was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific labeling was localized to the nuclei of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunohistochemistry
p70 S6 Kinase in Human Breast Cancer Tissue. p70 S6 Kinase was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human/Mouse/Rat p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of p70 S6 Kinase by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for p70 S6 Kinase at approximately 66 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human/Mouse/Rat p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
Background: p70 S6 Kinase
p70 S6 kinase is a Ser/Thr kinase activated by such mitogens as EGF, IGF-I, and insulin. The S6 protein of the 40S ribosomal subunit is a major substrate of p70 S6 kinase, and its phosphorylation upregulates the translation of mRNAs containing an oligopyrimidine tract at their transcriptional start site. The activity of p70 S6 kinase is controlled by multiple phosphorylation events, with phosphorylation at T229 by PDK1 and T389 by mTOR most critical for kinase function.
  • Entrez Gene IDs:
    6198 (Human); 72508 (Mouse); 83840 (Rat)
  • Alternate Names:
    EC 2.7.11; EC 2.7.11.1; p70 ribosomal S6 kinase alpha; p70 S6 kinase alpha; p70 S6 Kinase; p70 S6 kinase, alpha 1,70 kDa ribosomal protein S6 kinase 1; p70 S6 kinase, alpha 2; p70 S6KA; p70 S6K-alpha; p70(S6K)-alpha; p70-alpha; p70-S6K; P70S6K1; PS6K; ribosomal protein S6 kinase beta-1; Ribosomal protein S6 kinase I; ribosomal protein S6 kinase, 70kD, polypeptide 1; ribosomal protein S6 kinase, 70kDa, polypeptide 1; RPS6KB1; S6K; S6K1; S6K1p70-S6K 1; S6K-beta-1; serine/threonine kinase 14 alpha; Serine/threonine-protein kinase 14A; STK14A
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Applications
Sample Type
  1. Mild MPP(+) exposure-induced glucose starvation enhances autophagosome synthesis and impairs its degradation
    Authors: S Sakamoto, M Miyara, S Sanoh, S Ohta, Y Kotake
    Sci Rep, 2017;7(0):46668.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. The role of autophagy in cardiomyocytes in the basal state and in response to hemodynamic stress.
    Authors: Nakai A, Yamaguchi O, Takeda T
    Nat. Med., 2007;13(5):619-24.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
Cell and Tissue Staining Kits
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VisUCyte Antigen Retrieval Reagent-Basic

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Isotype Controls
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Normal Rabbit IgG Control

Ctrl AB-105-C 43  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
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Rabbit IgG HRP-conjugated Antibody

WB HAF008 23  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

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Rabbit IgG APC-conjugated Antibody

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Rabbit IgG PE-conjugated Antibody

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Rabbit IgG Fluorescein-conjugated Antibody

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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

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Goat Anti-Rabbit IgG Biotinylated Antibody

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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

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Rabbit IgG VisUCyte HRP Polymer Antibody

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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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