If you use 6-well or 12-well plates, total volume of the medium should be 3ml and 1.5ml, respectively, and decrease the amount of each reagent accordingly.2. One hour prior to the transfection, change to medium containing 25M chloroquine (from x1000 stock in PBS, stored at -20C). (Chloroquine can be omitted, but increases efficiency about x2) to a final volume of 500 l. Plaquenil and alcohol consumption Chloroquine phosphate powder Calcium Phosphate Transfection of 293T cells 2 Transfection 1. Twenty-four hours prior to transfection, inoculate 1 to 2 x106 cells/10 cm plate or, inoculate 3x106 cells/plate 16-hours prior to transfection. in 10 ml DMEM media + 10% BCS supplemented with L-glutamine and Pen/Strep. At the time of transfection, cells should be about 30-40%. Chloroquine phosphate tablets, Chloroquine phosphate, USP, is a 4-aminoquinoline compound for oral administration. It is a white, odorless, bitter tasting, crystalline substance, freely soluble inwater. Chloroquine phosphate tablets are an antimalarial and amebicidal drug. Chloroquine Phosphate *** The information contained here is subject to changes as I experiment and learn more about Chloroquine Phosphate *** What It Treats – Marine Ich Cryptocaryon irritans, Marine Velvet Disease Amyloodinium ocellatum, Brooklynella hostilis and Uronema marinum. How To. Add this mixture directly to the cells dropwise through the medium. As long as the cells are alive, they produce proteins. When you are ready, add 500 l of 2x HBS dropwise while gently mixing. Make sure you evenly sprinkle the droplet over the entire area.5. For secreted protein, you can change media once (at day 3 or 4, save the collected sup) and give it another 3-4 days of culture. Calcium phosphate transfection chloroquine Evaluation of Methods for Transient Transfection of a., Chloroquine - FDA prescribing information, side effects. Can you drink alcohol while on hydroxychloroquine Calcium Phosphate Transfection of adherent cells Reagents 2X HBS 280mM NaCl 8.18g/500ml pH with NaOH 10mM KCl 0.37g/500ml After adjusting volume with ddH Calcium Phosphate Transfection of adherent cells. Chloroquine phosphate REEF2REEF Saltwater and Reef Aquarium Forum. Calcium Phosphate Transfection - Thermo Fisher Scientific. The Calcium Phosphate Transfection Kit provides high-quality reagents to enable the introduction of DNA into eukaryotic cells via calcium phosphate co-precipitation. How it works The calcium phosphate transfection method for introducing DNA into mammalian cells is based on forming a calcium phosphate-DNA precipitate. Plate cells the night before to give 60-80% confluence at the day of transfection. 2. One hour prior to the transfection, change to medium containing 25µM chloroquine from x1000 stock in PBS, stored at -20°C. Volume should be 10 ml per dish. Chloroquine can be omitted, but increases efficiency about x2 3. This protocol is a modified version of a published method4, in which calcium phosphate–based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells.