p70S6K (pT389 + Total) ELISA Kit (ab225582)
Key features and details
- One-wash 90 minute protocol
- Sample type: Cell Lysate, Tissue Homogenate
- Detection method: Colorimetric
- Assay type: Semi-quantitative
- Reacts with: Mouse, Human
Overview
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Product name
p70S6K (pT389 + Total) ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% pT389 6 3.3% Total 6 4.5% Inter-assay Sample n Mean SD CV% pT389 3 2.9% Total 3 4.1% -
Sample type
Cell Lysate, Tissue Homogenate -
Assay type
Semi-quantitative -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Product overview
p70S6K (pT389 + Total) ELISA Kit (ab176665) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab225582). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying Total p70S6K protein in Human and mouse cells.
Abcam’s p70S6K (pT389) and p70S6K (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of p70S6K (pT389) and Total p70S6K 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.
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Notes
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Wash Buffer PT 1 x 15ml 50X Cell Extraction Enhancer Solution 1 x 1ml 5X Cell Extraction Buffer PTR 1 x 12ml Lyophilized p38 MAPK alpha Control Lysate 1 vial p70S6K (pT389) Capture Antibody 1 x 1.5ml p70S6K (pT389) Detector Antibody 1 x 1.5ml p70S6K (Total) Capture Antibody 1 x 1.5ml p70S6K (Total) Detector Antibody 1 x 1.5ml Plate Seal 1 unit SimpleStep Pre-Coated 96 Well Microplate (12 x 8 well strips) 1 unit Stop Solution 1 x 12ml TMB Substrate 1 x 12ml -
Research areas
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Cellular localization
P70 S6 Kinase: 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 -
Database links
- Entrez Gene: 6198 Human
- Entrez Gene: 6198 Human
- Entrez Gene: 6199 Human
- Entrez Gene: 58988 Mouse
- Entrez Gene: 72508 Mouse
- Entrez Gene: 72508 Mouse
- Entrez Gene: 361696 Rat
- Entrez Gene: 83840 Rat
see all
Images
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Other - p70S6K (pT389 + Total) ELISA Kit (ab225582)
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.
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Example of p70S6K (pT389) and p70S6K (Total) cell lysate standard curveBackground-subtracted data values (mean +/- SD) are graphed.
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Example of p70S6K (pT389) recombinant protein standard curveBackground-subtracted data values (mean +/- SD) are graphed.
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Cell line analysis for Total p70S6K from 100 µg/mL preparations of cell extractsData from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
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Inhibition of p70S6K (pT389) phosphorylation in MCF-7 cells in response to wortmannin treatmentMCF-7 cells were cultured in 96-well tissue culture plates, and treated (2 h) with a dose-range of wortmannin before stimulation with insulin (10 µg/mL for 30 min) and cell lysis. Data from triplicate measurements of p70S6K (pT389) are plotted and compared against total p70S6K protein levels.
