I wanna do haemagglutination assay, but I don't know what should i choose the RBC for this experiment.
I'm going to do animal experiments using mice and then I will analyze their antibody activity. Then am I have to use mouse RBC for HI test ? or Is it okay to use anythings?
And dose anyone elso who know how to make the RBC?
please answer me, thanks.
The RBC’s used in HA and HI assays are typically from chickens, turkeys, horses, guinea pigs, or humans depending on the selectivity of the targeted virus or bacterium and the associated surface receptors on the RBC
Preparation of human-O group or chicken RBC for HA test:
Red blood cell are often purchased from commercial sources, if you must bleed an animal, however, use 1 part of 5% sodium citrate to 5 parts of blood. Mix the blood-citrate solution by repeated inversions of tube. Then wash the cells 3 times in 0.85% saline: gently mix the citrate-blood suspension with 5 volumes of the saline. Sediment the suspension at 205g for 10 min, pour of the supernatant, add fresh saline, and repeat twice more. Drain of the last wash. Re-suspend the wash cell in saline to give a 10% stock solution. This may be kept at refrigerator temperature 4C for 5-6 days.
From this stock solution prepare a 0.5% suspension of RBC in PBS and use in your experiment
Chicken RBC and sheep RBC are the popular ones. Depending upon the relevance human RBC can be used.
Dear Garam Wi,
As far as I know the red blood cells of chicken and sheep are the most used. Best Regards
What is the amount of RBC per liter, to apply the hemglutination technique, thank you ..