Protein Name & Synonyms:
Serine/threonine-protein kinase STK11 (EC 22.214.171.124) (Liver kinase B1) (LKB1) (hLKB1) (Renal carcinoma antigen NY-REN-19)
- AMPK Signaling
- Insulin Signaling
This ELISA kit recognizes Human LKB1 phosphorylated at site Serine-428 as well as total LKB1.
Number of Targets Detected:
Compatible Sample Types:
- Cell Lysates
- Tissue Lysates
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
- 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 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
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 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.
COS cells were treated with Forskolin. Solubilize cells at 4 x 107
cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Forskolin Stimulation of COS Cell Line
COS cells were treated with Forskolin. Cell lysates were treated or untreated with Lambda Protein Phosphatase (LPP) and analyzed using this phosphoELISA and Western Blot.
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.