Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
R&D Systems | Catalog # AF8965
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Human, Mouse
Applications
Validated:
Western Blot, Simple Western
Cited:
Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
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Product Specifications
Immunogen
Phosphopeptide containing human, rat, and mouse p70 S6 Kinase T421/S424 sites
Specificity
Detects human, mouse and rat p70 S6 Kinase when phosphorylated at T421 and S424 and p85 S6K when phosphorylated at T444 and S447.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
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.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.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.Applications for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
Application
Recommended Usage
Simple Western
5 µg/mL
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1)
Sample: MCF‑7 human breast cancer cell line treated with Recombinant Human IGF‑I (Catalog # 291-G1)
Western Blot
0.25 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)
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)
Formulation, Preparation, and Storage
Purification
Antigen and protein A Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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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
Background: p70 S6 Kinase
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
- Ferrari, S. et al. (1994) Crit. Rev. Biochem. Mol. Biol. 29:385.
- Edelmann, H.M. et al. (1996) J. Biol. Chem. 271:963.
Alternate Names
p70-alpha, RPS6KB1, S6K1, STK14A
Gene Symbol
RPS6KB1
Additional p70 S6 Kinase Products
Product Documents for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
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.
Product Specific Notices for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
For research use only
Related Research Areas
Citations for Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
IL-7 Signaling Pathways
IL-9 Signaling Pathways
IL-15 Signaling Pathways
IL-21 Signaling Pathways
MAPK Signaling Pathway: Mitogen Stimulation Pathway
mTOR Signaling Pathway
TGF-beta Signaling Pathways