Description
Human/Mouse Phospho-Stat 1 (S727) and Total Stat 1 PharmaGenie ELISA - Kit Data
| Product SKU: | SBRS1981 |
| Size: | 96T |
| Application: | The antibody pair provided in this kit recognizes human and mouse Phospho-Stat1 (pSer727) + pan Stat1. |
| Uniprot: | P42224 |
| Gene ID: | 6772 |
| Gene Names: | STAT1 |
| Pathway: | JAK/STAT Signaling |
| Synonyms: | Signal transducer and activator of transcription 1-alpha/beta (Transcription factor ISGF-3 components p91/p84) |
| Target Species: | Human, Mouse |
| 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. |
Kit Components:
| 1. | Pre-Coated 96-well Strip Microplate |
| 2. | Wash Buffer |
| 3. | Anti-Phospho Antibody |
| 4. | Anti-Pan Antibody |
| 5. | HRP-Conjugated Secondary Antibody |
| 6. | Streptavidin-Conjugated HRP |
| 7. | Assay Diluent |
| 8. | TMB One-Step Substrate |
| 9. | Stop Solution |
| 10. | Lysis Buffer |
| 11. | Positive Control Sample |
Other materials and equipment required:
The Assay Genie Human/Mouse Phospho-Stat 1 (S727) and Total Stat 1 ELISA Kit 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
Kit Protocol:
| 1. | Prepare all reagents and samples as instructed in the manual. |
| 2. | Add 100 ul of sample or positive control to each well. |
| 3. | Incubate 2.5 h at RT or O/N at 4 °C. |
| 4. | Add 100 ul of prepared primary antibody to each well. |
| 5. | Incubate 1 h at RT. |
| 6. | Add 100 ul of prepared 1X HRP-Streptavidin to each well. |
| 7. | Incubate 1 h at RT. |
| 8. | Add 100 ul of TMB One-Step Substrate Reagent to each well. |
| 9. | Incubate 30 min at RT. |
| 10. | Add 50 ul of Stop Solution to each well. |
| 11. | Read at 450 nm immediately. |
