Overview

  • Product name

    p70S6K (pT389) ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    MCF7 extract 6 3.3%
    Inter-assay
    Sample n Mean SD CV%
    MCF7 extract 3 2.9%
  • Sample type

    Cell Lysate, Tissue Homogenate
  • Assay type

    Semi-quantitative
  • Sensitivity

    1 ng/ml
  • Range

    1 ng/ml - 100 ng/ml
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    Abcam’s p70S6K (pT389) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of p70S6K (pT389) protein in Human and mouse cells.

    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    p70S6K (pT389) Capture Antibody 1 x 3ml
    p70S6K (pT389) Detector Antibody 1 x 3ml
    10X Wash Buffer PT 1 x 15ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Lyophilized p70S6K Control Lysate 1 vial
    Plate Seal 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • Research areas

  • Relevance

    The protein p70 S6 kinase is critical for cell cycle progression and cell survival. In response to mitogen stimulation, p70 S6 kinase activation up-regulates ribosomal biosynthesis and enhances the translational capacity. The p70S6K phosphorylates the S6 protein of the 40S subunit of the ribosome. This kinase was first characterized as an insulin/mitogen-activated protein kinase, whose major known substrate is the 40 S ribosomal subunit protein S6. The p70 S6 kinase is activated by diverse stimuli through a multi-site phosphorylation such as Thr-252 and Ser-434. In Alzheimer's Disease (AD), p70 S6 kinase activation is associated with PHF-tau (hyperphosphorylated tau) accumulation. In non-neuronal cells, p70 S6 kinase have been shown to regulate actin polymerization.
  • Cellular localization

    Cell junction › synapse › synaptosome By similarity. Mitochondrion outer membrane. Mitochondrion. Note: Colocalizes with URI1 at mitochondrion. Isoform Alpha I: Nucleus. Cytoplasm Isoform Alpha II: Cytoplasm
  • Alternative names

    • 70 kDa ribosomal protein S6 kinase 1
    • p70 alpha
    • p70 S6 kinase alpha
    • p70 S6K
    • p70 S6KA
    • p70(S6K) alpha
    • p70s6kA
    • PS6K
    • Ribosomal protein S6 kinase 70kDa polypeptide 1
    • ribosomal protein S6 kinase beta 1
    • ribosomal protein S6 kinase I
    • Ribosomal protein S6 kinase, 70-kD
    • RPS6KB1
    • S6K
    • S6K1
    • Serine/threonine kinase 14 alpha
    • Serine/threonine protein kinase 14A
    • STK14A
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab176651 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Example of a typical p70S6K (pT389) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Example of a typical p70S6K (pT389) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of p70S6K (pT389) are normalized and plotted.

  • Cell line analysis for Total p70S6K from 100 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

  • Induction of p70S6K (pT389) phosphorylation in MCF-7 cells in response to insulin treatment. MCF-7 cells were cultured in 96-well tissue culture plates, serum-starved and treated (10 min) with a dose-range of insulin before cell lysis. Data from quadruplicate measurements of p70S6K (pT389) are plotted and compared against total p70S6K protein levels. Comparative p70S6K (pT389) and p70S6K (Total) data also shown by Western Blot.

Protocols

References

This product has been referenced in:

  • Zhang X  et al. Downregulation of NLRP2 inhibits HUVEC viability by inhibiting the MAPK signaling pathway. Mol Med Rep N/A:N/A (2018). Read more (PubMed: 30431084) »
See 1 Publication for this product

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