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AKT2 Western Blot Kit
AKT2 Western Chemiluminescent Blotting Kit
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
| Host | Rat |
|---|---|
| Conjugate | Unconjugated |
| Target Species | Human |
| Reactivity | Human |
| Clonality | Monoclonal |
Application 
| WB, I, LCI |
| Application Note | Use Rockland Immunochemicals’ Anti-AKT2 Chemiluminescent Kit for Western Blotting for detection of human AKT2 native and recombinant proteins by western blot. Expect a band approximately 56 kDa in size corresponding to AKT2 protein. This kit is useful for both western blotting and dot blotting methods. Please read the entire product insert prior to use. |
| Preservative | Wash buffers MUST NOT contain SODIUM AZIDE or other inhibitors of peroxidase activity! |
| Gene ID | 208 |
|---|---|
| Purity | The kit is designed to detect both unphosphorylated and phosphorylated forms of the protein. Cross reactivity with AKT2 from other species has not been determined, however, the sequence of the immunogen shows 100% identity to human, mouse, and rat, therefore, cross reactivity is expected. Cross-reactivity with AKT1 and AKT3 has not been determined. This kit contains sufficient substrate for up to 5 mini blots at 7.5 x 8 cm2 (1,800 cm2) and is stable for at least 1 year when stored as indicated. |
| Storage Condition | See kit insert for complete instructions. |
| Precautions Note | This product is for research use only and is not intended for therapeutic or diagnostic applications. |
| Name | AKT2 (HGNC:392) |
|---|---|
| Function | Serine/threonine kinase closely related to AKT1 and AKT3. All 3 enzymes, AKT1, AKT2 and AKT3, are collectively known as AKT kinase. AKT regulates many processes including metabolism, proliferation, cell survival, growth and angiogenesis, through the phosphorylation of a range of downstream substrates. Over 100 substrates have been reported so far, although for most of them, the precise AKT kinase catalyzing the reaction was not specified. AKT regulates glucose uptake by mediating insulin-induced translocation of the SLC2A4/GLUT4 glucose transporter to the cell surface. Phosphorylation of PTPN1 at 'Ser-50' negatively modulates its phosphatase activity preventing dephosphorylation of the insulin receptor and the attenuation of insulin signaling. Phosphorylation of TBC1D4 triggers the binding of this effector to inhibitory 14-3-3 proteins, which is required for insulin-stimulated glucose transport. AKT also regulates the storage of glucose in the form of glycogen by phosphorylating GSK3A at 'Ser-21' and GSK3B at 'Ser-9', resulting in inhibition of its kinase activity. Phosphorylation of GSK3 isoforms by AKT is also thought to be one mechanism by which cell proliferation is driven. AKT regulates also cell survival via the phosphorylation of MAP3K5 (apoptosis signal- related kinase). Phosphorylation of 'Ser-83' decreases MAP3K5 kinase activity stimulated by oxidative stress and thereby prevents apoptosis. AKT mediates insulin-stimulated protein synthesis by phosphorylating TSC2 at 'Ser-939' and 'Thr-1462', thereby activating mTORC1 signaling and leading to both phosphorylation of 4E-BP1 and in activation of RPS6KB1. AKT is involved in the phosphorylation of members of the FOXO factors (Forkhead family of transcription factors), leading to binding of 14-3-3 proteins and cytoplasmic localization. In particular, FOXO1 is phosphorylated at 'Thr-24', 'Ser-256' and 'Ser-319'. FOXO3 and FOXO4 are phosphorylated on equivalent sites. AKT has an important role in the regulation of NF-kappa-B-dependent gene transcription and positively regulates the activity of CREB1 (cyclic AMP (cAMP)-response element binding protein). The phosphorylation of CREB1 induces the binding of accessory proteins that are necessary for the transcription of pro-survival genes such as BCL2 and MCL1. AKT phosphorylates 'Ser- 454' on ATP citrate lyase (ACLY), thereby potentially regulating ACLY activity and fatty acid synthesis. Activates the 3B isoform of cyclic nucleotide phosphodiesterase (PDE3B) via phosphorylation of 'Ser-273', resulting in reduced cyclic AMP levels and inhibition of lipolysis. Phosphorylates PIKFYVE on 'Ser-318', which results in increased PI(3)P- 5 activity. The Rho GTPase-activating protein DLC1 is another substrate and its phosphorylation is implicated in the regulation cell proliferation and cell growth. AKT plays a role as key modulator of the AKT-mTOR signaling pathway controlling the tempo of the process of newborn neurons integration during adult neurogenesis, including correct neuron positioning, dendritic development and synapse formation. Signals downstream of phosphatidylinositol 3-kinase (PI(3)K) to mediate the effects of various growth factors such as platelet- derived growth factor (PDGF), epidermal growth factor (EGF), insulin and insulin-like growth factor I (IGF-I). AKT mediates the antiapoptotic effects of IGF-I. Essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. May be involved in the regulation of the placental development (PubMed:21432781, PubMed:21620960). In response to lysophosphatidic acid stimulation, inhibits the ciliogenesis cascade. In this context, phosphorylates WDR44, hence stabilizing its interaction with Rab11 and preventing the formation of the ciliogenic Rab11-FIP3-RAB3IP complex. Also phosphorylates RAB3IP/Rabin8, thus may affect RAB3IP guanine nucleotide exchange factor (GEF) activity toward Rab8, which is important for cilia growth (PubMed:31204173). |
| Cellular Location | Cytoplasm. Nucleus. Cell membrane; Peripheral membrane protein. Early endosome {ECO:0000250|UniProtKB:Q60823}. Note=Through binding of the N-terminal PH domain to phosphatidylinositol (3,4,5)- trisphosphate (PtdIns(3,4,5)P3) or phosphatidylinositol (3,4)- bisphosphate (PtdIns(3,4)P2), recruited to the plasma membrane. Cell membrane recruitment is facilitated by interaction with CLIP3 Colocalizes with WDFY2 in early endosomes (By similarity). Localizes within both nucleus and cytoplasm in proliferative primary myoblasts and mostly within the nucleus of differentiated primary myoblasts (PubMed:17565718). {ECO:0000250|UniProtKB:Q60823, ECO:0000269|PubMed:17565718} |
| Tissue Location | Widely expressed. Expressed in myoblasts (PubMed:17565718). |
Research Areas
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Application Protocols
Provided below are standard protocols that you may find useful for product applications.
Background
Rockland Immunochemicals’ Chemiluminescent Western Blot Kit for AKT2 combines all of the necessary reagents with a rapid proven protocol and the extremely high signal detection of our luminol chemiluminescent substrate for the detection of native and/or recombinant human AKT2 protein. AKT2 Western Blotting kit allows for the detection of endogenous protein levels of human AKT2 present in cell lysates provided by the user. As a positive control, this kit includes a MDA-MB468 whole cell lysate proven to contain AKT2. After protein separation by SDS-PAGE and transfer, the membrane is probed with Rockland's optimized Anti-AKT2 antibody. Detection of the membrane bound antibody-antigen complex is achieved by the addition of a secondary antibody conjugated to the enzyme horseradish peroxidase. The enzyme reacts with a specialized formulation of luminol, an extremely sensitive, non-radioactive substrate that emits light and allows visualization using X-ray film or other imaging methods, including highly sensitive CCD cameras and imaging systems. AKT2 Western Blotting is ideal for investigators involved in Cell Signaling, Cancer, Neuroscience and Signal Transduction research.
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$ 385.00
Cat# ASR5944
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