phospho-p70 S6 Kinase (T421/S424) Antibody

Name: phospho-p70 S6 Kinase (T421/S424) Antibody
Brand: R&D Systems
SKU: AF8965
Price: 169 USD
Availability: InStock

R&D Systems | Catalog # AF8965

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Citations (7)

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

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

View all p70 S6 Kinase Primary Antibodies »

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 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 Western^TM.

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 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)

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)

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.

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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).