Human p70 S6 Kinase Antibody

R&D Systems | Catalog # AF8962

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Simple Western, CyTOF-ready

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all p70 S6 Kinase Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human p70 S6K
Accession # M60725

Specificity

Detects human 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.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human p70 S6 Kinase Antibody

Detection of Human and Mouse p70 S6 Kinase antibody by Western Blot.

Detection of Human p70 S6 Kinase by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (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.
Detection of p70 S6 Kinase antibody in HeLa Human Cell Line antibody 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 p70 S6 Kinase Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8962, filled histogram) or control antibody (AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
p70 S6 Kinase antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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 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; 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.
p70 S6 Kinase antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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 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; 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 antibody by Simple WesternTM.

Detection of p70 S6 Kinase by Simple WesternTM.

Simple Western lane view shows lysates of MCF-7 human breast cancer 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 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.
Detection of Human p70 S6 Kinase/S6K by Western Blot

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 p70 S6 Kinase Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunohistochemistry

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

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with methanol

Simple Western

2 µg/mL
Sample: MCF-7 human breast cancer cell line

Western Blot

0.2 µg/mL
Sample: MCF-7 human breast cancer cell line, and HeLa human cervical epithelial carcinoma cell line

Reviewed Applications

Read 3 reviews rated 5 using AF8962 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

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. See Certificate of Analysis for details.<br/>*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: 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.

Alternate Names

p70-alpha, RPS6KB1, S6K1, STK14A

Entrez Gene IDs

6198 (Human); 72508 (Mouse); 83840 (Rat)

Gene Symbol

RPS6KB1

UniProt

M60725

Additional p70 S6 Kinase Products

Product Documents for Human p70 S6 Kinase 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.

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Product Specific Notices for Human p70 S6 Kinase Antibody

For research use only

Citations for Human p70 S6 Kinase Antibody

Customer Reviews for Human p70 S6 Kinase Antibody (3)

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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human/Mouse/Rat p70 S6 Kinase Antibody
    Name: Khuynh Nguyen
    Application: Western Blot
    Sample Tested: MDA-MB-231 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 04/18/2022
  • Human/Mouse/Rat p70 S6 Kinase Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Adipose tissue
    Species: Mouse
    Verified Customer | Posted 10/23/2019
    Human p70 S6 Kinase Antibody AF8962
  • Human/Mouse/Rat p70 S6 Kinase Antibody
    Name: Joseph Moore
    Application: Western Blot
    Sample Tested: Cartilage tissue
    Species: Mouse
    Verified Customer | Posted 08/14/2019
    Dilution 1:100 overnight incubation, we used the P70 as a baseline for Phospho-P70
    Human p70 S6 Kinase Antibody AF8962

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Protocols

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