title: Phospho-specific antibody for cell signaling proteins,specially for kinases and their targets
status: Public on Jun 17 2026
contact.city: Vancouver
contact.first: Xiaoyan
contact.last: Jiang
contact.email: anny_chen@hotmail.com
contact.state: BC
contact.address: 675 w10th Ave
contact.department: TFL
contact.country: Canada
contact.institute: BC Cancer Research Center
contact.phone: 6048757411
organism: Mus musculus
manufacture_protocol: The Kinex KAM-1325 antibody microarray - Cells were lysed in homogenizing buffer with ice-cold Tris (2-carboxyethyl)phosphosine (TCEP), within 10 minutes 6 mM 2-nitro-5-thiocyanatobenzoic acid (NTCB) was added, and the samples sonicated for 40 seconds in intervals of 10 seconds with 10 seconds intermissions over ice. The resulting homogenates were further incubated for 30 minutes during ultracentrifugation for at 20°C at 50,000 RPM. Approximately 100 µg (protein) of each samples were then labeled with 20 μg of NHS-Biotin (APExBIO) for 1 hour. The resulting biotin-labeled protein fragments were then purified using Sephadex G-25 Spin columns (Sigma-Aldrich) and eluted. These samples (~100 µg) were then diluted to 400 μL with phosphate-buffered saline (PBS) with 0.05% Tween and 50 mM ethanolamine, and then applied to the antibody microarray slides and incubated in a humidity chamber for 2 hours. Slides were then washed, and anti-biotin antibody labelled with Alex546/Cy3 dye was then applied over the slides for 10 minutes in the humidity chamber. The slides were then washed, dried under nitrogen, and scanned at 543 nM using a ScanArray GX Microarray scanner (Perkin-Elmer).