HRP-conjugated Phospho-AKT (Ser473) Monoclonal antibody HRP-66444 proteintech
$479.00
In stock
SKU
HRP-66444
| Catalog Number | HRP-66444 |
| Synonyms | AKT (Ser473), p AKT, p AKT (Ser473), p AKT Ser473, p-AKT |
| Host | Mouse |
| Reactivity | human, mouse |
| Form | Liquid |
| Formulation | PBS, Proclin300, BSA, Glycerol |
| Applications | WB |
| Host / Isotype | Mouse / IgG1 |
| Tested Applications — Positive WB detected in | Calyculin A treated HEK-293 cells |
| Recommended Dilutions — Western Blot (WB) | WB : 1:5000-1:50000 |
| Positive WB detected in | Calyculin A treated HEK-293 cells |
| Western Blot (WB) | WB : 1:5000-1:50000 |
| Tested Reactivity | human, mouse |
| Cited Reactivity | human, mouse |
| Class | Monoclonal |
| Type | Antibody |
| Immunogen | Peptide Predict reactive species |
| Full Name | v-akt murine thymoma viral oncogene homolog 1 |
| Observed Molecular Weight | 60-62 kDa |
| GenBank Accession Number | NM_005163 |
| Gene Symbol | AKT1 |
| Gene ID (NCBI) | 207 |
| RRID | AB_3083816 |
| Conjugate | HRP |
| Purification Method | Protein A purification |
| UNIPROT ID | P31749 |
| Storage Buffer | PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3. |
| Storage Conditions | Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20 o C storage. |
| WB protocol for HRP Phospho-AKT (Ser473) antibody HRP-66444 | Download protocol |
| human,mouse | WB Chem Biodivers UPLC-MS/MS-Network Pharmacology and Experimental Validation Reveal Active Components of Ficus tikoua for Ameliorating Liver Fibrosis. Authors - Shi-Ying Xu View Article |
| Citations | 1 |
| Dilutions | WB : 1:5000-1:50000 |
| Isotype | IgG1 |
| Clonality | Monoclonal |
| Clone Number | 1C10B8 |
| Conjugation | HRP |
1) What is AKT? The serine/threonine kinase B AKT pathway (also known as the PI3K-Akt pathway) plays a vital role in the regulation of cellular processes, including cell proliferation, survival, and growth - processes that are essential for oncogenesis. Mutation of the regulator proteins PI3K and PTEN causes uncontrolled disruption within the PI3-kinase pathway, leading to the development of human cancers (1,2; see also AKT pathway poster for more details). 2) phospho-AKT and FAQs A) What is the best way to normalize phosphorylated proteins analyzed by western blot? Normalize phospho-AKT and total AKT with your loading control (e.g. Actin, tubulin), then calculate the phospho/total ratio using these normalized values. Put more simply: 1. Calculate the ratio of band intensities of a phospho-AKT band: the loading control. 2. Calculate the ratio of band intensities of total AKT: loading control. 3. Divide ratio obtained #1 by #2 to obtain a normalized value for comparison among different conditions. This procedure allows one to distinguish between a change in AKT expression and a change in the ratio of phospho-AKT. * If you are looking at the differences in a phospho-AKT expression resulting from an experimental condition (e.g., knockdown), you should also show the expression of total AKT to distinguish between a change in AKT expression (transcription/translation level) and a change in the AKT phosphorylation status. B) What is the observed molecular weight for AKT and phospho-AKT? Molecular Weight AKT - 56 kDa Molecular Weight phospho-AKT - 60 kDa (Figure 1) Figure 1. WB: HEK-293 cell lysate was subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT antibody) and 66444-1-Ig (AKT-phospho-S473 antibody) at a dilution of 1:4000 incubated at room temperature for 1.5 hours. C) Are there any special WB conditions to optimize staining of a phospho-AKT? Since this is a phosphorylated protein, 5% BSA is recommended over non-fat milk as a blocking agent. D) What are good positive and negative controls for a phospho-AKT? - Positive Control: HEK293 cells - Negative Control: Treatment with PI3K inhibitors (e.g. wortmannin) E) What species does this antibody react with? Our internal testing has confirmed that it reacts with the human and mouse forms of phospho-AKT.Reactivity with the human form is also supported by the literature's citations of this antibody . References: 1. Perturbations of the AKT signaling pathway in human cancer. 2. Targeting the PI3K-Akt pathway in human cancer: rationale and promise. Protocols Product Specific Protocols WB protocol for HRP Phospho-AKT (Ser473) antibody HRP-66444 Download protocol Standard Protocols Click here to view our Standard Protocols
Publications
- UPLC-MS/MS-Network Pharmacology and Experimental Validation Reveal Active Components of Ficus tikoua for Ameliorating Liver Fibrosis.
Journal: Chem Biodivers | Authors: Shi-Ying Xu | Species: human,mouse | Application: WB
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