AB253298

AKT 1/2/3 Total ELISA Kit

SimpleStep
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AKT 1/2/3 Total ELISA Kit is a single-wash 90-min Simplestep used to quantify AKT 1/2/3 Total with a sensitivity of 1 µg/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

View Alternative Names

PKBG, AKT3, RAC-gamma serine/threonine-protein kinase, Protein kinase Akt-3, Protein kinase B gamma, RAC-PK-gamma, STK-2, PKB gamma

4 Images

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

sELISA

Unknown

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Example of a typical AKT1 Total recombinant protein standard curve.

Background-subtracted data values (mean +/- SD) are graphed.

sELISA

Unknown

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Example of a typical AKT 1/2/3 Total control lysate dilution series in 1X Cell Extraction Buffer PTR.

Raw data values are shown in the table.

sELISA

Unknown

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Induction of AKT 1/2/3 (pS473) phosphorylation in MCF-7 cells in response to insulin treatment, and levels of AKT 1/2/3 Total in the same samples.

MCF-7 cells were cultured in 96-well tissue culture plates, serum-starved (2 hr) and treated (30 min) with a dose-range of insulin before cell lysis. Data from duplicate measurements of AKT 1/2/3 (pS473) are plotted and compared against AKT 1/2/3 Total protein levels.

sELISA

Unknown

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Cell line analysis for AKT 1/2/3 Total from preparations of 17,500 cells/well cell extracts.

Data from triplicate measurements (mean +/- SD) are plotted.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Reacts with

Mouse, Human

Assay type

Semi-quantitative

Sensitivity

< 1 µg/mL

Range

0.05 - 3.12 %

Assay time

1h 30m

Assay Platform

Microplate

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Reactivity data

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

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Product details

AKT 1/2/3 Total (ab253298) is a single-wash 90 min sandwich ELISA designed for the semi-quantitative measurement of total AKT 1/2/3 protein in human and mouse cell lysates. It uses our proprietary SimpleStep ELISA® technology. Semi-quantitate total AKT 1/2/3 with 1 μg/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 SimpeStep ELISA® kits.

AKT isoforms belong to a conserved family of serine/threonine protein kinases. AKT plays a central role in cell survival and inhibiting apoptosis, by phosphorylating and inactivating several targets. AKT is activated by insulin and various other agonists, which promote transport to the plasma membrane, where the activation loop of AKT is phosphorylated at Thr308 by PDK1. The carboxy terminus is phosphorylated at Ser473 by mammalian target of rapamycin complex 2 (TORC2).

AKT substrates include Bad, forkhead transcription factors, and caspase-9. Another vital AKT function is the regulation of glycogen synthesis through phosphorylation of GSK-3. AKT phosphorylation also prevents GSK-3β mediated phosphorylation and degradation of cyclin D1, and negatively regulates the cyclin dependent kinase inhibitors p27 Kip and p21 Waf1/CIP1. AKT also plays a critical role in the regulation of TORC1 and tuberin (TSC2).

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": "Overall", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "9.5" }, { "reproducibilityType": "Intra", "sample": "Overall", "replicates": 6, "mean": null, "standardDeviation": null, "coefficientOfVariability": "1.8" } ]

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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab253298-1x96Tests", "assetComponentDetails": [ { "size":"1 x 3 mL", "name":"AKT 1/2/3 Total Capture Antibody", "number":"AB253298-CMP03", "productcode":"" }, { "size":"1 x 15 mL", "name":"10X Wash Buffer PT", "number":"AB253298-CMP02", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated 96-Well Microplate (ab206978)", "number":"AB253298-CMP01", "productcode":"" }, { "size":"1 x 3 mL", "name":"AKT 1/2/3 Total Detector Antibody", "number":"AB253298-CMP04", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Lyophilized AKT Control Lysate", "number":"AB253298-CMP06", "productcode":"" }, { "size":"1 x 12 mL", "name":"TMB Substrate", "number":"AB253298-CMP09", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution", "number":"AB253298-CMP05", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seal", "number":"AB253298-CMP07", "productcode":"" }, { "size":"1 x 12 mL", "name":"5X Cell Extraction Buffer PTR", "number":"AB253298-CMP10", "productcode":"" }, { "size":"1 x 12 mL", "name":"Stop Solution", "number":"AB253298-CMP08", "productcode":"" } ] } } }

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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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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

AKT proteins also known as protein kinase B (PKB) include AKT1 AKT2 and AKT3 playing key roles in cellular processes. These serine/threonine kinases have molecular masses ranging between 55 to 60 kDa. AKT protein family expresses widely in multiple tissues including muscle liver and brain. Each isoform has specific but overlapping roles in cell signaling. AKT kinases often studied with ELISA kits facilitate insight into essential cellular regulation mechanisms.

Biological function summary

AKT proteins drive processes like cell survival growth and proliferation. They interact with various proteins forming complexes that regulate pathways contributing to these cellular functions. AKT activation occurs in response to signals from growth factor receptors leading to downstream effects such as protein synthesis and glycogen metabolism. These proteins also exhibit regulatory influence in apoptosis contributing to their extensive role in maintaining cellular integrity.

Pathways

AKT proteins integrate into major signaling networks. The PI3K-AKT pathway is a critical route that bounces signals from the cell membrane to regulate cell survival and growth. AKT interacts with mTOR an important component of cell growth and metabolism pathways establishing a cross-talk between them. This convergence serves as a regulatory hub orchestrating numerous downstream effects affecting cellular functions.

AKT proteins have notable implications in cancer and diabetes. Dysregulation of AKT activity associates with oncogenesis through alterations in cell proliferation and survival pathways. Moreover AKT2 plays a significant role in glucose metabolism connecting it to insulin signaling and diabetes development. These proteins also interact with PTEN a tumor suppressor affecting pathways important for understanding their roles in disease pathology.

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Product protocols

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Target data

AKT3 is one of 3 closely related serine/threonine-protein kinases (AKT1, AKT2 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT3 is the least studied AKT isoform. It plays an important role in brain development and is crucial for the viability of malignant glioma cells. AKT3 isoform may also be the key molecule in up-regulation and down-regulation of MMP13 via IL13. Required for the coordination of mitochondrial biogenesis with growth factor-induced increases in cellular energy demands. Down-regulation by RNA interference reduces the expression of the phosphorylated form of BAD, resulting in the induction of caspase-dependent apoptosis.

See full target information AKT3

Additional targets

RAC-beta serine/threonine-protein kinase,AKT3

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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AKT 1/2/3 Total ELISA Kit

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Sandwich ELISA - AKT 1/2/3 Total ELISA Kit (AB253298)

Key facts

Detection method

Sample types

Reacts with

Assay type

Sensitivity

Range

Assay time

Assay Platform

Reactivity data

Product details

Precision

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Additional targets

Complementary Products

Assay Kits

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Product promise