The RayBiotech Phospho-AKT (S473) ELISA Kit measures phosphorylated AKT (Ser473) and total (pan) AKT in cell and tissue lysates.
Phospho-AKT (S473) ELISA Kit
Phospho-AKT (S473) ELISA Kit
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species | Human, Mouse, Rat |
| Specificity | The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473). |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| Accession Number | P31749 |
| Gene Id | 391 |
| Gene Symbols | AKT1, PKB, RAC |
| Protein Name / Synonyms | RAC-alpha serine/threonine-protein kinase (EC 2.7.11.1) (Protein kinase B) (PKB) (Protein kinase B alpha) (PKB alpha) (Proto-oncogene c-Akt) (RAC-PK-alpha) |
| Quantitative/Semi-Quantitative | Semi-Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Angiogenesis, Apoptosis, Post-Translational Modifications, Inflammation, Phosphorylation, mTOR Signaling, PI3K-AKT Signaling, Insulin Signaling, Autophagy, Akt Signaling |
| Shipping Type | Blue ice |
| Storage | -20°C |
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Product Features
- Simultaneously measure phosphorylated protein and pan protein in one experiment (for normalization purpose)
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- Anti-Pan Antibody (*not included in phospho-only version)
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
Other Materials Required
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Benchtop rocker or shaker
- Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Storage/Stability
Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.
documents
citations
Icosapent ethyl alleviates acetic acid-induced ulcerative colitis via modulation of SIRT1 signaling pathway in rats
- Species:
- Rat
- Sample Type:
- colon
- Year:
- 2022
- Journal Name:
- International Immunopharmacology
IL-10 directly protects cortical neurons by activating PI-3 kinase and STAT-3 pathways
- Species:
- Rat
- Sample Type:
- Cell Lysate (Primary Rat neuronal cells)
- Year:
- 2011
- Journal Name:
- Brain Research
