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Human/Mouse/Rat Phospho-PDK1 (Ser410) Cell-Based ELISA Kit
RayBio® Human/Mouse/Rat Phospho-PDK1 (Ser410) Cell-Based ELISAt. This assay semi-quantitatively measures PDK1 phosphorylated at Serine-410 in adherent cell lines.
Product Description
Specifications
| Size |
1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species |
Human, Mouse, Rat |
| Protein Name / Synonyms |
3-phosphoinositide-dependent protein kinase 1 (hPDK1) (EC 2.7.11.1) |
| Specificity |
This ELISA kit recognizes phosphorylated human, mouse or rat PDK1 (S410) in adherent whole cell lines. |
| Accession Number |
O15530|Q9Z2A0|O55173 |
| Gene Symbols |
PDK1|PDPK1 |
| Gene Id |
5170 |
| Quantitative/Semi-Quantitative |
Semi-Quantitative |
| Compatible Sample Types |
Adherent Cell Culture |
| Solid Support |
96-well Microplate |
| Method Of Detection |
Colorimetric |
| Design Principle |
Cell-based |
| Research Area |
Post-Translational Modifications, Phosphorylation |
| Estimated Lead Time |
1-2 business days |
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- Rapidly measure phosphorylated protein in adherent cell lines
- Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
- No sample lysis is needed
- Compatible with a standard ELISA plate reader
Kit Components
- Uncoated 96-well Strip Microplate
- Wash Buffers
- Fixing Solution
- Quenching Buffer
- Blocking Buffer
- Anti-phospho antibody
- Anti-pan antibody
- HRP-Conjugated Secondary Antibody
- TMB One-Step Substrate
- Stop 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
Protocol Outline
- Seed 10,000-30,000 cells into each well and incubate overnight.
- Apply various treatment, inhibitors or activators according to manufacturer'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.
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.
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