Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody

Catalog # Availability Size / Price Qty
AF8965
AF8965-SP
Detection of Human and Mouse Phospho-p70 S6 Kinase (T421/S424) by Western Blot.
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Citations (6)
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Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody Summary

Species Reactivity
Human, Mouse, Rat
Specificity
Detects human, mouse and rat p70 S6 Kinase when phosphorylated at T421 and S424 and p85 S6K when phosphorylated at T444 and S447.
Source
Polyclonal Rabbit IgG
Purification
Antigen and protein A Affinity-purified
Immunogen
Phosphopeptide containing human, rat, and mouse p70 S6 Kinase T421/S424 sites
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.25 µg/mL
See below
Simple Western
5 µg/mL
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.

Scientific Data

Western Blot Detection of Human and Mouse Phospho-p70 S6 Kinase (T421/S424) antibody by Western Blot. View Larger

Detection of Human and Mouse Phospho-p70 S6 Kinase (T421/S424) by Western Blot. Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL of Recombinant Human IGF-1 (Catalog # 291-G1) for 20 minutes and NIH-3T3 mouse embryonic fibroblast cell line untreated or treated with 100 ng/mL of Human PDGF (Catalog # 120-HD) for 20 minutes. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8965), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-p70 S6 Kinase (T421/S424) at approximately 70 and 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western Detection of Human Phospho-p70 S6 Kinase (T421/S424) antibody by Simple Western<SUP>TM</SUP>. View Larger

Detection of Human Phospho-p70 S6 Kinase (T421/S424) 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 0.2 mg/mL. A specific band was detected for Phospho-p70 S6 Kinase (T421/S424) at approximately 66 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8965). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Western Blot Detection of Human p70 S6 Kinase/S6K by Western Blot View Larger

Detection of Human p70 S6 Kinase/S6K by Western Blot Time-dependent changes in autophagic induction with mild MPP+ exposure.(a) SH-SY5Y cells were exposed to 10 and 200 μM MPP+ for up to 48 h. Time-dependent changes in the phosphorylation levels of various proteins were estimated by western blotting. (b,c,d) SH-SY5Y cells were exposed to 10 and 200 μM MPP+ for 24 h (b), 36 h (c) or 48 h (d) with or without 200 nM bafilomycin A1 (Baf) for the last 4 h; LC3-II turnover was estimated by western blotting at various time points. (e,f,g) SH-SY5Y cells were exposed to 10 and 200 μM MPP+ for 24 h (e), 36 h (f) or 48 h (g) and subsequently immunostained with an anti-Atg16L antibody. The numbers of Atg16L-positive puncta per cell were evaluated at various time points. Data are expressed as means ± S.D. from at least three independent experiments. **p < 0.01. Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/srep46668), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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.

Background: p70 S6 Kinase

p70 S6 Kinase (p70S6K) is responsible for the phosphorylation of 40S ribosomal protein S6 and is ubiquitously expressed in human adult tissues (1). p70S6K is activated by serum stimulation and this activation is inhibited by wortmannin and rapamycin. p70S6K activity undergoes changes in the cell cycle and increases
20-fold in G1 cells released from G0 (2). p70S6K activation requires sequential phosphorylations at proline-directed residues in the putative autoinhibitory pseudosubstrate domain, as well as T389, a site phosphorylated by phosphoinositide-dependent kinase 1 (PDK1).

References
  1. Ferrari, S. et al. (1994) Crit. Rev. Biochem. Mol. Biol. 29:385.
  2. Edelmann, H.M. et al. (1996) J. Biol. Chem. 271:963.
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

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Citations for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody

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.

6 Citations: Showing 1 - 6
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  1. IER family proteins are regulators of protein phosphatase PP2A and modulate the phosphorylation status of CDC25A
    Authors: T Ueda, Y Kohama, H Sakurai
    Cell. Signal., 2018-12-30;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. IL-4 upregulates cyclooxygenase-1 expression in macrophages
    Authors: AE Shay, BT Diwakar, BJ Guan, V Narayan, JF Urban, KS Prabhu
    J. Biol. Chem., 2017-07-06;0(0):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. 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, 2017-04-26;7(0):46668.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  4. Progesterone receptor membrane component 1/Sigma-2 receptor associates with MAP1LC3B and promotes autophagy.
    Authors: Mir, Shakeel, Schwarze, Steven R, Jin, Ling, Zhang, Jinling, Friend, Woodrow, Miriyala, Sumitra, St Clair, Daret, Craven, Rolf J
    Autophagy, 2013-09-04;9(10):1566-78.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Impaired Pten expression in human malignant peripheral nerve sheath tumours.
    Authors: Bradtmoller M, Hartmann C, Zietsch J, Jaschke S, Mautner V, Kurtz A, Park S, Baier M, Harder A, Reuss D, von Deimling A, Heppner F, Holtkamp N
    PLoS ONE, 2012-11-06;7(11):e47595.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  6. 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., 2007-04-22;13(5):619-24.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot

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