RBC Lysis/Fixation Solution (10X)

Buffers/Solutions
Sony
Flow Cytometry
50 ml
2712005
$146.00

Description

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.

Formulation

This RBC Lysis/Fixation Solution is supplied as a 10X solution and should be diluted to 1X in room temperature deionized water.

Recommended Usage


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.

References

1. Horvath P, et al. 2012. J Infect Dis. PubMed
2. Kusner LL, et al. 2014. PLoS One. 9:102231. PubMed