In order to better help customers to provide real primary cells, immunofluorescence is used to identify the extracted cell types. Immunofluorescence technology, also known as fluorescent antibody technology, is one of the earliest developments in marker immunology. It is a technology built on the basis of immunology, biochemistry and microscopy. The main features of this technology are: high specificity, high sensitivity and fast speed. The following is the step of immunofluorescent cell identification provided by the Cypress technicians, for reference only:
Cell climber
Four pieces of glass were taken in a 24-well plate, and 1 mL of the medium was added to each well, and 0.02 ml/well was added to the cells. Place the incubator for 2h or overnight.
2. Fixed
After the cells were climbed, the medium was aspirated, washed once with PBS, and fixed with 4% PFA for 30 min at 4 °C. Wash with PBS for 3 x 5 min/time. It is also possible not to aspirate the PBS for the last time and let it sit at 4 ° C overnight.
3. Broken film closure
Remove the moisture from the slide and place it on the petri dish support. Place the glass slide solution: 0.5% Trition X-100 and PBS 1:1, add 10% serum, take 50uL membrane to seal the droplets on the waterproof membrane. , cover the cell with one side of the slide for 2 hours.
4. Primary antibody incubation <br> Primary antibody configuration: After diluting the antibody with PBS 1:100 (200), take 50uL of primary antibody on the waterproof membrane (in the wet box), and cover the cell with one side of the slide. At 4 ° C (up to one week)
The negative control group can replace the primary antibody with PBS.
5. Secondary antibody incubation
Wash PBS 3 × 5min / time,
Secondary anti-mixture configuration: secondary antibody: PBS=1:500
DAPI: PBS=1:1000
A 50 uL secondary antibody mixture was dropped on the waterproof membrane, covered with a slide, and protected from light, and allowed to stand at room temperature for 2 hours.
6. Embedding
Wash PBS for 3 × 5 min / time, drop 1 drop of Fluoromount-G on each slide, and cover one side of the cells.
Alcohol Fructose Syrup,High Fructose Corn Syrup Juice,Fructose In Agave Syrup,Glucose Fructose Syrup Dangers
JILIN COFCO BIO-CHEM AND BIO-ENERGY MARKETING CO., LTD , https://www.cofco-biotech.com