Description
Human Phospho-MLKL (S345) and Total MLKL PharmaGenie ELISA Kit
| Product SKU: | SBRS1901 |
| Size: | 96T |
| Application: | The antibody pair provided in this kit recognizes Human MLKL phosphorylated at site Serine-345 as well as total MLKL. |
| Uniprot: | Q8NB16 |
| Gene ID: | 197259 |
| Pathway: | Necroptosis |
| Synonyms: | Mixed lineage kinase domain-like protein (hMLKL) |
| Target Species: | Human |
| 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. |
- 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 Phospho-MLKL (S345) and Total MLKL PharmaGenie ELISA Kit (SBRS1901) 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.
 | HT29 cells were treated with Calyculin A. Cell lysates were analyzed using this phosphoELISA and Western Blot. |
 | HT29 cells were treated with Calyculin A. Solubilize cells at 4 x 10^7 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. |
