Description
Human/Mouse/Rat Phospho-AKT (S473) PharmaGenie ELISA Kit
| Product SKU: | SBRS1743 |
| Size: | 96T |
| Application: | The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Akt (pSer473). |
| Uniprot: | P31749 |
| Gene ID: | 391 |
| Gene Names: | AKT1 / PKB / RAC |
| Pathway: | Insulin Signaling / mTOR Signaling / PI3K-AKT Signaling |
| 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) |
| Target Species: | Human, Mouse, Rat |
| Compatible Sample Types: | Cell Lysates, Tissue Lysates |
| Design Principle: | Sandwich-based |
| Method of Detection: | Colorimetric |
| Quantitative/Semi-Quantitative: | Semi-Quantitative |
| 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. |
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
Other materials and equipment required:
The Assay Genie Human/Mouse/Rat Phospho-AKT (S473) PharmaGenie ELISA Kit (SBRS1743) will require other equipment and materials to carry out the assay. Please see list below for further details.
- 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 ul 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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 ul of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 ul of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 ul of prepared 1X HRP-Streptavidin to each well.
- Incubate 1 h at RT.
- Add 100 ul of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 ul of Stop Solution to each well.
- Read at 450 nm immediately.
