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AKT1 + AKT2 + AKT3 (pS473) ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the semi-quantitative measurement of AKT1 + AKT2 + AKT3 (pS473) Mouse, Human in Tissue Homogenate, Cell Lysate samples.

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Images

Key facts

Detection method

Colorimetric

Sample types

Tissue Homogenate, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mouse, Human

Range

1 - 100 ng/mL

Assay time

1h 30m

Sensitivity

= 30 pg/mL

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Reactivity data

Application

ELISA

Reactivity

Reacts

Dilution info

-

Notes

-

Target data

Function

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.

Alternative names

What's included?

1 x 96 Tests
Components
10X Wash Buffer PT
1 x 15 mL
50X Cell Extraction Enhancer Solution
1 x 1 mL
5X Cell Extraction Buffer PTR
1 x 12 mL
AKT 1/2/3 (pS473) Capture Antibody
1 x 3 mL
AKT 1/2/3 (pS473) Detector Antibody
1 x 3 mL
Lyophilized AKT Control Lysate
1 x 1 Vial
Plate Seal
1 x 1 Unit
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Substrate
1 x 12 mL

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AKT1 + AKT2 + AKT3 (pS473) ELISA Kit is a single-wash 90-min SimpleStep ELISA® for the semi-quantitative measurement of AKT1 + AKT2 + AKT3 (pS473) Mouse, Human in Tissue Homogenate, Cell Lysate samples.

Alternative names

Key facts

Detection method

Colorimetric

Sample types

Tissue Homogenate, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mouse, Human

Range

1 - 100 ng/mL

Assay time

1h 30m

Assay Platform

Microplate

Sensitivity

= 30 pg/mL

General recovery

4.2%

Precision

Intra assay

Sample

HEK-293 Extract

n

6

mean

-

SD

-

C.V.

= 5.4

Inter assay

Sample

HEK-293 Extract

n

3

mean

-

SD

-

C.V.

= 2.7

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

Abcam's AKT 1/2/3 (pS473) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of AKT 1/2/3 (pS473) 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.

As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.

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.

Supplementary info

Biological function summary

AKT proteins regulate cell cycle growing cell survival proliferation and metabolism. They participate as core components of the PI3K/AKT/mTOR signaling pathway forming complexes with other proteins to transmit signals. They bind to phosphoinositide lipids on the cell membrane facilitating their activation and downstream signaling. Through these activities the AKT transporter proteins maintain cellular homeostasis and play a part in stress response.

Activity summary

AKT1 AKT2 and AKT3 also known as Protein Kinase B (PKB) isoforms are serine/threonine-specific protein kinases with critical roles in cellular processes. AKT1 has a molecular weight of about 55.8 kDa AKT2 weighs approximately 56.1 kDa and AKT3 typically has a similar mass. These proteins are expressed in many tissues including brain and heart with AKT1 ubiquitously present AKT2 focused in insulin-responsive tissues and AKT3 mainly in the brain. The molecular weight of AKT plays an important role in their functionality and specificity in tissues.

Pathways

AKT proteins engage in important signaling networks including the PI3K/AKT pathway and mTOR pathway. They work closely with PI3K and mTOR proteins coordinating cellular growth and energy metabolism. In particular the AKT pathway responds to growth factors and insulin influencing glucose uptake and glycolysis regulation through interaction with proteins such as glycogen synthase kinase 3 (GSK3).

Associated diseases and disorders

Dysregulation of AKT signaling can lead to cancer and diabetes. High AKT activation correlates with various cancers by promoting cell survival and growth. In diabetes impaired AKT2 regulation disrupts glucose uptake affecting insulin response. AKT's relationship with mTOR is significant as it often influences tumor growth and progression in cancerous tissues.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635), expandable thumbnail

    ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635)

  • Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635), expandable thumbnail

    Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635)

    Typical cell lysate dilution series.

    Example of a typical AKT (pS473) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635), expandable thumbnail

    Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635)

    Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of AKT1 (pS473) are normalized and plotted.

  • Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635), expandable thumbnail

    Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635)

    AKT (pS473) phosphorylation in response to insulin treatment.

    Induction of AKT (pS473) 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 (5 min) with a dose-range of insulin before cell lysis. Data from quadruplicate measurements of AKT (pS473) are plotted and compared against total AKT1 protein levels. Comparative AKT (pS473) and AKT1 (Total) data also shown by Western Blot.

  • Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635), expandable thumbnail

    Sandwich ELISA - AKT1 + AKT2 + AKT3 (pS473) ELISA Kit (ab176635)

    Comparison of total AKT1 expression in different cell lines.

    Cell line analysis for total AKT1 from 20 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

Downloads

Product protocols

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

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

For licensing inquiries, please contact partnerships@abcam.com

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