Description
Human/Mouse/Rat Phospho-MKK6 (S207) PharmaGenie ELISA Kit and Total MKK6 PharmaGenie ELISA Kit (SBRS1899)
Product SKU: | SBRS1899 |
Size: | 96T |
Application: | The antibody pair provided in this kit recognizes human, mouse, and rat MKK6 phosphorylated at site Serine-207 as well as total MKK6. |
Uniprot: | P52564 |
Gene ID: | 5608 |
Gene Names: | MAP2K6 / MEK6 / MKK6 / PRKMK6 / SKK3 |
Pathway: | MAPK Signaling |
Synonyms: | Dual specificity mitogen-activated protein kinase kinase 6 (MAP kinase kinase 6) (MAPKK 6) (EC 2.7.12.2) (MAPK/ERK kinase 6) (MEK 6) (Stress-activated protein kinase kinase 3) (SAPK kinase 3) (SAPKK-3) (SAPKK3) |
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), 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. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C. |
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- Anti-Pan Antibody
- HRP-Conjugated Secondary Antibody
- Streptavidin-Conjugated HRP
- 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-MKK6 (S207) PharmaGenie ELISA Kit and Total MKK6 PharmaGenie ELISA Kit (SBRS1899) 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.
HeLa cells were treated or untreated with UV. Cell lysates were analyzed using this phosphoELISA and Western Blot. | |
HeLa cells were treated with UV. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. |