Protein Name & Synonyms:
Mitogen-activated protein kinase 8 (MAP kinase 8) (MAPK 8) (EC 220.127.116.11) (JNK-46) (Stress-activated protein kinase 1c) (SAPK1c) (Stress-activated protein kinase JNK1) (c-Jun N-terminal kinase 1)
The antibodies provided in this kit recognizes human, mouse and rat JNK phosphorylated at sites Thr183 and Tyr185 as well as total JNK for comparison.
Number of Targets Detected:
Method of Detection:
1, 2, or 5 x 96-Well Microplate Kit
- Site and signal pathway-specific
- In vitro detection of adherent cell culture
- No sample lysis is needed
- Compatible with a standard ELISA plate reader
- Faster results than with ELISA
- Adaptable for high-throughput screening and drug discovery
Kit Componentsuncoated 96-well Microplate Wash Buffer AWash Buffer BFixing SolutionQuenching BufferBlocking BufferAnti-phospho antibodyAnti-pan antibodyHRP-Conjugated Secondary AntibodyTMB One-Step SubstrateStop Solution
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
- Seed 10,000-30,000 cells into each well and incubate overnight.
- Apply various treatment, inhibitors or activators according to manufacture's instructions.
- Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
- Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
- Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
- Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
- Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 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.
The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.