pS351/pS353-S1P1 Phosphorylation Assay Kit

Supplied Reagents:
• 20 mL Detergent Buffer (Cell Lysis Buffer) for 96-well plates
• 2 x Protease and Phosphatase Inhibitor Tablets (dissolve 1 tablet into 10 mL Detergent Buffer before use)
• 8.0 mL Anti-HA Magnetic Bead Solution
• 0.6 mL Premium Phosphosite-Specific Antibody for detection of phosphorylated receptors
• 0.6 mL Premium Non-Phospho Antibody for detection of total receptors
• 1.2 mL HRP-Labelled Secondary Antibody for detection
• 20 mL Detection Solution
• 20 mL Stop Solution

Required Materials:
• Handheld magnetic separation block (e.g. cat. #VP 771HH-H, V&P Scientific) or automated microplate washer equipped with a magnet (e.g. Biotek 405 LS)
• Microplate centrifuge (e.g. Thermo Fisher Multifuge X Pro, 4,000 x g, 4°C)
• Multichannel pipette
• Orbital microplate shaker (e.g. Corning LSE digital microplate shaker)
• Microplate reader for measuring OD at 405 nm
• Cell culture plates: poly-D-lysine (PDL)-coated 96-well plates, F-bottom (e.g. cat. #655940, Greiner Bio-One)
• Assay plates: 96-well plates, U-bottom, transparent or white (e.g. cat. #650101, Greiner Bio-One)
• Detection plates, 96-well plates, F-bottom, transparent, (e.g. cat. #655101, Greiner Bio-One)
• Phosphate-buffered saline containing Ca2+/Mg2+ (PBS) and PBS with 0.1% Tween 20 (PBST)