| 存储条件 |
|---|
| Product Name | Phospho-AKT (S473) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser473 of human Akt1. |
| Modification | p-S473 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCIHCWB |
| WB:1:2000-1:5000 IHC:1:200 FC:1:10000 | |
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | AKT1 |
|---|---|
| Gene Synonyms | AKT PKB RAC PRKBA PKB-ALPHA RAC-ALPHA |
| Gene Full Name | AKT serine/threonine kinase 1 |
| Gene Summary | This gene encodes one of the three members of the human AKT serine-threonine protein kinase family which are often referred to as protein kinase B alpha, beta, and gamma. These highly similar AKT proteins all have an N-terminal pleckstrin homology domain, a serine/threonine-specific kinase domain and a C-terminal regulatory domain. These proteins are phosphorylated by phosphoinositide 3-kinase (PI3K). AKT/PI3K forms a key component of many signalling pathways that involve the binding of membrane-bound ligands such as receptor tyrosine kinases, G-protein coupled receptors, and integrin-linked kinase. These AKT proteins therefore regulate a wide variety of cellular functions including cell proliferation, survival, metabolism, and angiogenesis in both normal and malignant cells. AKT proteins are recruited to the cell membrane by phosphatidylinositol 3,4,5-trisphosphate (PIP3) after phosphorylation of phosphatidylinositol 4,5-bisphosphate (PIP2) by PI3K. Subsequent phosphorylation of both threonine residue 308 and serine residue 473 is required for full activation of the AKT1 protein encoded by this gene. Phosphorylation of additional residues also occurs, for example, in response to insulin growth factor-1 and epidermal growth factor. Protein phosphatases act as negative regulators of AKT proteins by dephosphorylating AKT or PIP3. The PI3K/AKT signalling pathway is crucial for tumor cell survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating AKT1 which then phosphorylates and inactivates components of the apoptotic machinery. AKT proteins also participate in the mammalian target of rapamycin (mTOR) signalling pathway which controls the assembly of the eukaryotic translation initiation factor 4F (eIF4E) complex and this pathway, in addition to responding to extracellular signals from growth factors and cytokines, is disregulated in many cancers. Mutations in this gene are associated with multiple types of cancer and excessive tissue growth including Proteus syndrome and Cowden syndrome 6, and breast, colorectal, and ovarian cancers. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jul 2020] |
| Alternative Names | AKT1 antibody AKT1_HUMAN antibody MGC99656 antibody PKB antibody PKB-ALPHA antibody PRKBA antibody Protein Kinase B Alpha antibody Protein kinase B antibody Proto-oncogene c-Akt antibody RAC Alpha antibody AKT1 antibody AKT1_HUMAN antibody MGC99656 antibody PKB antibody PKB-ALPHA antibody PRKBA antibody Protein Kinase B Alpha antibody Protein kinase B antibody Proto-oncogene c-Akt antibody RAC Alpha antibody RAC antibody RAC-alpha serine/threonine-protein kinase antibody RAC-PK-alpha antibody akt s473 antibody |
| Molecular Weight(MW) | 56kDa |
| Cellular Localization | Cytoplasm, Nucleus, Cell membrane. |
WB
Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody at 1/2,000 dilution and pan AKT antibody at 1/2,000 dilution. Lane 1: MCF7 cell lysate, Lane 2: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, Lane 3: SH-SY5Y cell lysate, Lane 4: SH-SY5Y treated with 100ng/mL PDGF for 5 minutes cell lysate, Lane 5: HEK-293 cell lysate, Lane 6: HEK-293 treated with 50μM LY294002 for 6 hours cell lysate, Lane 7: NIH/3T3 cell lysate, Lane 8: NIH/3T3 treated with 100ng/mL PDGF for 5 minutes cell lysate, Lane 9: C6 cell lysate, Lane 10: C6 treated with 100ng/mL PDGF for 5 minutes cell lysate, Lane 11: MCF7 treated with 50ng/mL Calyculin A for 45 minutes cell lysate, then the membrane treated with λpp for 1 hour, Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA for 1 hour at room temperature. The primary antibody at 1/2,000 dilution and pan AKT antibody at 1/2,000 dilution were used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Phospho-AKT (S473) antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IF
Flow cytometric analysis of NIH/3T3 cells untreated (left) / treated with 100ng/mL PDGF for 1 hour (right) labeling Phospho-AKT (S473). Cells were fixed and permeabilized. Then stained with the primary antibody (0.1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
WB
(a–c) ECA109 and KYSE150 cells were treated each day with 5 μM GO-203 or CP-2 for 3 days, and GO-203 inhibits the phosphorylation of AKT and S6K1 protein and then increases PDCD4 leave in ESCC cells.| Application Notes | WB:1:2000-1:5000 IHC:1:200 FC:1:10000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
抱歉，暂无相关文献
