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- Sony Biotechnology
- Sony Biotechnology
RBC Lysis/Fixation Solution has been designed and optimized for one-step lysing of red blood cells (RBCs) and fixation of the remaining blood elements (such as leukocytes) following immunofluorescent staining with fluorochrome-conjugated antibodies. It can be used for either lyse/wash or lyse/no wash immunofluorescent staining procedures. This solution is supplied as a 10X concentrate. Dilute to 1X working solution with room temperature dI H20 prior to use.
Whole Blood Lyse/Wash Procedure:
1. Prepare 1X RBC Lysis/Fixation solution by diluting 1 part of RBC Lysis/Fixation solution with 9 parts of room temperature dI H20.
2. Mix 100 microL of whole blood with fluorochrome-conjugated antibody.
3. Incubate at room temperature, in the dark, for 20 minutes.
4. Add 2.0 mL 1X RBC Lysis/Fixation solution, gently mixing each tube immediately.
5. Incubate at room temperature, in the dark, for 15 minutes.
6. Centrifuge at 350Xg for five minutes. Discard supernatant.
7. Add 2 mL Cell Staining Buffer (Cat No 420201).
8. Centrifuge at 350Xg for five minutes. Discard supernatant.
9. Resuspend cell pellet in 0.3-0.5 mL Cell Staining Buffer.
10. Analyze cells immediately by flow cytometry or store at 4°C, in the dark, for no longer than 12 hours.
Whole Blood Lyse/No Wash Procedure:
1. Prepare 1X RBC Lysis/Fixation solution by diluting 1 part of RBC Lysis/Fixation solution with 9 parts room temperature dI H20.
2. Mix 50 microL of whole blood with fluorochrome-conjugated antibody.
3. Incubate at room temperature, in the dark, for 20 minutes.
4. Add 0.45 mL 1X RBC Lysis/Fixation solution, gently mixing each tube immediately.
5. Incubate at room temperature, in the dark, for 15 minutes.
6. Analyze samples by flow cytometry.