Human AKT3 (pS473) ELISA Kit (ab270887)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 19.6 U/ml
- Range: 46.88 U/ml - 3000 U/ml
- Sample type: Cell Lysate
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human AKT3 (pS473) ELISA Kit
See all AKT3 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Extract 3 2.5% Inter-assay Sample n Mean SD CV% Extract 24 3.4% -
Sample type
Cell Lysate -
Assay type
Sandwich (quantitative) -
Sensitivity
19.6 U/ml -
Range
46.88 U/ml - 3000 U/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell Lysate 97 94% - 107% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human ATK (pS473) ELISA kit (ab270887) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of ATK (pS473) protein in human cell extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate human ATK (pS473) with 5.9 units/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 stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
AKT3 is a member of the AKT family that includes AKT1 and AKT2. These serine/threonine-protein kinases are involved in diverse processes including glucose uptake, cell cycle progression, and apoptosis. AKT3 plays an important role in brain development and is crucial for glioma cells malignancy. Phosphorylation of Serine 473 is a known regulator of AKT’s kinase activity. The antibodies for this kit were raised against an AKT3-specific N-terminal fragment, as well as a phospho-peptide encompassing the 473-serine site, which shares high sequence identity for all three AKT members. The protein standard in this product is recombinant human AKT3 ligated with peptide containing phosphorylated Serine 416.
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
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human AKT3 (pS473) Capture Antibody 1 x 600µl 10X Human AKT3 (pS473) Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human AKT3 (pS473) Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
IGF-1 leads to the activation of AKT3, which may play a role in regulating cell survival. Capable of phosphorylating several known proteins. Truncated isoform 2/PKB gamma 1 without the second serine phosphorylation site could still be stimulated but to a lesser extent. -
Tissue specificity
In adult tissues, it is highly expressed in brain, lung and kidney, but weakly in heart, testis and liver. In fetal tissues, it is highly expressed in heart, liver and brain and not at all in kidney. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 PH domain.
Contains 1 protein kinase domain. -
Domain
Binding of the PH domain to the phosphatidylinositol 3-kinase alpha (PI(3)K) results in its targeting to the plasma membrane. -
Post-translational
modificationsPhosphorylation on Thr-305 and Ser-472 is required for full activity (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
Ubiquitinated. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome. -
Cellular localization
Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. - Information by UniProt
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Alternative names
- AKT
- Akt3
- AKT3 kinase
see all -
Database links
- Entrez Gene: 10000 Human
- Omim: 611223 Human
- SwissProt: Q9Y243 Human
- Unigene: 498292 Human
Images
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SimpleStep ELISA Protocol Diagram
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 human AKT3 (pS473) standard curve in 1X Cell Extraction Buffer PTR.The AKT3 (pS473) 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.
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Interpolated concentrations of native AKT3 (pS473) in human SH-SY5Y +pervanadate extract 500 µg/mL extract load.The concentrations of AKT3 (pS473) were measured in duplicate and interpolated from the AKT3 (pS473) standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean AKT3 (pS473) concentration was determined to be 2,820 units/mL.
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Comparison of native AKT3 (pS473) concentration in treated versus control cell extracts.SH-SY5Y cells were treated with 10 mM pervanadate for 20 minutes, scraped and buffer exchanged into PBS + 20 mM NaF, followed by protein extraction. Mock control SH-SY5Y cells accompanied cell treatment. Concentrations of AKT3 (pS473) were measured in duplicate and interpolated against the standard curve. Results show the mean AKT3 (pS473) concentration in control cells was 30.6 units/mL and 2,795 units/mL in pervanadate treated samples.
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Sandwich ELISA - Human AKT3 (pS473) ELISA Kit (ab270887)To learn more about the advantages of recombinant antibodies see here.
