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AB283535

Human p70S6K ELISA Kit

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Human p70S6K ELISA Kit is a single-wash 90-min Simplestep used to quantify Human p70S6K with a sensitivity of 21.5 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

View Alternative Names

STK14A, RPS6KB1, Ribosomal protein S6 kinase beta-1, S6K-beta-1, S6K1, 70 kDa ribosomal protein S6 kinase 1, Ribosomal protein S6 kinase I, Serine/threonine-protein kinase 14A, p70 ribosomal S6 kinase alpha, P70S6K1, p70-S6K 1, p70 S6 kinase alpha, p70 S6K-alpha, p70 S6KA

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Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)
  • sELISA

Supplier Data

Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)

Example of human p70S6K standard curve in 1X Cell Extraction Buffer PTR.

The p70S6K standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)
  • sELISA

Supplier Data

Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)

Interpolated concentrations of native p70S6K in human MCF7 cell starved for 24 hours in media without serum then treated with or without 100 ng/mL human IGF-I for 20 minutes based on 8 μg/mL extracts.

The concentrations of p70S6K were measured in duplicate and interpolated from the p70S6K standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean p70S6K concentration was determined to be 5,525 pg/mL in IGF-I treated MCF7 extract and 6,131 pg/mL in untreated MCF7 extract. Note that interpolated p70S6K1 concentrations are not largely affected by condition that induce the target phosphorylation.

Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)
  • sELISA

Supplier Data

Sandwich ELISA - Human p70S6K ELISA Kit (AB283535)

Interpolated concentrations of native p70S6K in mouse C2C12 cell based on 200 µg/mL extract load.

The concentrations of p70S6K were measured in duplicate and interpolated from the human p70S6K standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean p70S6K concentration was determined to be 4,395 pg/mL in mouse C2C12 extract.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Human

Assay type

Sandwich (quantitative)

Sensitivity

= 21.5 pg/mL

Range

140.63 - 9000 pg/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human p70S6K ELISA kit (ab272199) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human p70S6K protein in human cell and tissue extract samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human p70S6K with 21.5 pg/mL sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

P70S6K, also known as Ribosomal protein S6 kinase beta-1, is a serine/threonine-protein kinase that acts downstream of mTOR signaling in response to growth factors and nutrients to promote cell proliferation, cell growth and cell cycle progression. P70S6K activation requires multiple phosphorylation events on serine/threonine residues. The activation appears to be first mediated by phosphorylation of multiple sites in the autoinhibitory domain, which facilitates phosphorylation at Thr-412, disrupting the autoinhibitory mechanism and allowing phosphorylation of Thr-252 by PDPK1. The active conformation of the kinase is believed to be stabilized by a mechanism involving three conserved phosphorylation sites located in the kinase domain activation loop (Thr-252) and in the AGC-kinase C-terminal domain (Ser-394 in the middle of the tail/linker region and Thr-412 within a hydrophobic motif at its end). P70S6K protein is highly homologous between the species. Relatively to the human protein, Macaca mulatta p70S6K is 100% identical, mouse p70S6K 99.4%, rat p70S6K 99.6%, and cow p70S6K 99.8% identical.

The standard is affinity purified.

REACH authorisation
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.

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Precision

[ { "reproducibilityType": "Inter", "sample": "Extract", "replicates": 0, "mean": null, "standardDeviation": null, "coefficientOfVariability": null }, { "reproducibilityType": "Intra", "sample": "Extract", "replicates": 10, "mean": null, "standardDeviation": null, "coefficientOfVariability": "2.5" } ]
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Recovery

[ { "sample": "Cell Lysate", "range": "96 - 98 %", "average": "= 97" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

Serine/threonine-protein kinase that acts downstream of mTOR signaling in response to growth factors and nutrients to promote cell proliferation, cell growth and cell cycle progression (PubMed : 11500364, PubMed : 12801526, PubMed : 14673156, PubMed : 15071500, PubMed : 15341740, PubMed : 16286006, PubMed : 17052453, PubMed : 17053147, PubMed : 17936702, PubMed : 18952604, PubMed : 19085255, PubMed : 19720745, PubMed : 19935711, PubMed : 19995915, PubMed : 22017876, PubMed : 23429703, PubMed : 28178239). Regulates protein synthesis through phosphorylation of EIF4B, RPS6 and EEF2K, and contributes to cell survival by repressing the pro-apoptotic function of BAD (PubMed : 11500364, PubMed : 12801526, PubMed : 14673156, PubMed : 15071500, PubMed : 15341740, PubMed : 16286006, PubMed : 17052453, PubMed : 17053147, PubMed : 17936702, PubMed : 18952604, PubMed : 19085255, PubMed : 19720745, PubMed : 19935711, PubMed : 19995915, PubMed : 22017876, PubMed : 23429703, PubMed : 28178239). Under conditions of nutrient depletion, the inactive form associates with the EIF3 translation initiation complex (PubMed : 16286006). Upon mitogenic stimulation, phosphorylation by the mechanistic target of rapamycin complex 1 (mTORC1) leads to dissociation from the EIF3 complex and activation (PubMed : 16286006). The active form then phosphorylates and activates several substrates in the pre-initiation complex, including the EIF2B complex and the cap-binding complex component EIF4B (PubMed : 16286006). Also controls translation initiation by phosphorylating a negative regulator of EIF4A, PDCD4, targeting it for ubiquitination and subsequent proteolysis (PubMed : 17053147). Promotes initiation of the pioneer round of protein synthesis by phosphorylating POLDIP3/SKAR (PubMed : 15341740). In response to IGF1, activates translation elongation by phosphorylating EEF2 kinase (EEF2K), which leads to its inhibition and thus activation of EEF2 (PubMed : 11500364). Also plays a role in feedback regulation of mTORC2 by mTORC1 by phosphorylating RICTOR, resulting in the inhibition of mTORC2 and AKT1 signaling (PubMed : 19720745, PubMed : 19935711, PubMed : 19995915). Also involved in feedback regulation of mTORC1 and mTORC2 by phosphorylating DEPTOR (PubMed : 22017876). Mediates cell survival by phosphorylating the pro-apoptotic protein BAD and suppressing its pro-apoptotic function (By similarity). Phosphorylates mitochondrial URI1 leading to dissociation of a URI1-PPP1CC complex (PubMed : 17936702). The free mitochondrial PPP1CC can then dephosphorylate RPS6KB1 at Thr-412, which is proposed to be a negative feedback mechanism for the RPS6KB1 anti-apoptotic function (PubMed : 17936702). Mediates TNF-alpha-induced insulin resistance by phosphorylating IRS1 at multiple serine residues, resulting in accelerated degradation of IRS1 (PubMed : 18952604). In cells lacking functional TSC1-2 complex, constitutively phosphorylates and inhibits GSK3B (PubMed : 17052453). May be involved in cytoskeletal rearrangement through binding to neurabin (By similarity). Phosphorylates and activates the pyrimidine biosynthesis enzyme CAD, downstream of MTOR (PubMed : 23429703). Following activation by mTORC1, phosphorylates EPRS and thereby plays a key role in fatty acid uptake by adipocytes and also most probably in interferon-gamma-induced translation inhibition (PubMed : 28178239).
See full target information RPS6KB1 phospho T389
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