Protein Name & Synonyms:
Cytoplasmic tyrosine-protein kinase BMX (EC 18.104.22.168) (Bone marrow tyrosine kinase gene in chromosome X protein) (Epithelial and endothelial tyrosine kinase) (ETK) (NTK38)
The antibody pair provided in this kit recognizes human tyrosine-phosphorylated BMX
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
- Biotinylated Anti-Phosphotyrosine Antibody
- Stop Solution
- Assay Diluent(s)
- Positive Control Sample
- Lysis Buffer
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
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.
HUVEC cells were treated with Pervanadate (PV). Cells were solubilzed at 4 x 107
cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA (see Reagent Preparation step 4).
PV Stimulation of HUVEC Cell Line
HUVEC cells were untreated or treated with PV. Cell lysates were 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.