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Product Details
| Cat # +Size | K435-40 |
|---|---|
| Size | 40 assays |
| Detection Method | Western blot analysis using anti-Phospho-GSK-3α (Ser 21) antibody at 1:1000 dilutions |
| Species Reactivity | Mammalian |
| Applications | The Akt Activity Assay Kit utilizes an Akt-specific antibody to immunoprecipitate Akt from cell lysate. Activity of the Akt is then determined in a kinase reaction using recombinant GSK-3α as substrate. |
| Features & Benefits | • Simple procedure • The kit is suitable for both research and drug discovery • The assay utilizes a N-terminal c-Jun fusion protein bead to selectively “pull down” JNK from cell lysate c-Jun phosphorylation is then measured using a phospho-c-Jun specific antibody. |
| Kit Components | • Kinase Extraction Buffer • Akt Specific Antibody • Protein A Sepharose • GSK-3α Protein/ATP Mixture • Kinase Assay Buffer • Phospho-GSK-3α Specific Antibody |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Akt is a protein kinase that can be activated by insulin and various growth factors and functions in a pathway involving PI3 kinase. Recent evidence suggests that Akt functions to promote cell survival by actively inhibiting apoptosis. The Akt Activity Assay Kit utilizes an Akt-specific antibody to immunoprecipitate Akt from cell lysate. Activity of the Akt is then determined in a kinase reaction using recombinant GSK-3α as substrate. Phosphorylation of the GSK-3α can be analyzed by Western blot analysis using the phospho-GSK-3α specific antibody included in the kit. The kit specifically detects Akt1, Akt2, and Akt3 activities, other kinase activities would not be detected.
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What is the proper protocol for tissue extracts?
10-100mg of tissue can be homogenized using the extraction buffer and spun down to get the tissue homogenate.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Yuanfu Xu et al., Cigarette smoke (CS) and nicotine delay neutrophil spontaneous death via suppressing production of diphosphoinositol pentakisphosphate. PNAS, May 2013; 110: 7726 - 7731.
Yuanfu Xu et al., Cigarette smoke (CS) and nicotine delay neutrophil spontaneous death via suppressing production of diphosphoinositol pentakisphosphate. PNAS, Apr 2013; 10.1073/pnas.1302906110.
Ching-Chieh Su et al., Mitochondrial Akt-regulated mitochondrial apoptosis signaling in cardiac muscle cells, Am J Physiol Heart Circ Physiol, Feb 2012; 302: H716 - H723.
Braydich-Stolle, L. et al. Silver Nanoparticles Disrupt GDNF/Fyn Kinase Signaling in Spermatogonial Stem Cells. Toxicol. Sci., 2010; 116: 577-589.
Wei Y et al. (2007) Mol. Pharmacol. 71: 835-842.
For more citations of this product click here
