The FIX & PERM Cell Permeabilization Kit protocol does not say what volume my cells should be in when they are added to the tube. What should I do?
Your cells should be in approximately 100 µL volume. One way to do this is to aliquot the correct number of cells (10E6) and then spin them down and decant the supernatant. The residual volume should be approximately 100 µL.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can I use the FIX & PERM Cell Permeabilization Kit to access nuclear epitopes?
Yes. Generally, you need to increase the incubation time in the permeabilization buffer (Reagent B). Here are a couple of references:
-Millard I, Degrave E, Philippe M et al. (1998) Detection of intracellular antigens by flow cytometry: comparison of two chemical methods and microwave heating. Clin Chem 44:2320-2330(http://www.ncbi.nlm.nih.gov/pubmed/9799760).
-Parker JE, Mufti GJ, Rasool F et al. (2000) The role of apoptosis, proliferation, and the Bcl-2-related proteins in the myelodysplastic syndromes and acute myeloid leukemia secondary to MDS. Blood 96:3932-3938(http://www.ncbi.nlm.nih.gov/pubmed/11090080).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The FIX & PERM Cell Permeabilization Kit is used to prepare cells for flow cytometry; can I use it for immunohistochemistry?
Yes. Here are a couple of references:
Haranaga S, Yamaguchi H, Friedman H et al.(2001) Chlamydia pneumoniae infects and multiplies in lymphocytes In vitro. Infect Immun 69:7753-7759 (http://www.ncbi.nlm.nih.gov/pubmed/11705957).
Al-Mufti R, Hambley H, Albaiges G et al. (2000) Increased fetal erythroblasts in women who subsequently develop pre-eclampsia. Hum Reprod 15:1624-1628 (http://www.ncbi.nlm.nih.gov/pubmed/10875878).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Does the FIX & PERM Cell Permeabilization Kit (Cat. Nos. GAS003 and GAS004) lyse red blood cells?
Yes. The FIX & PERM Cell Permeabilization Kit is used in flow cytometry for fixing and permeabilizing cells in suspension, allowing access of antibody to intracellular targets without sacrificing scatter characteristics. It has been validated for use in cell lines, whole blood, and bone marrow. In addition fixing and permeabilizing cells, it lyses red blood cells. It is labeled as a general purpose reagent (GPR) and can be used in clinical settings, but it is also used widely in research labs.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
