Frequently asked questions about using eBioscience flow cytometry

Q. Can I vortex an antibody or recombinant protein?

    We do not recommend oscillating proteins or antibody solutions, which can alter the characteristics of the product. It is usually possible that the liquid has already been dispersed in the pipe during transportation. It is recommended to centrifuge immediately before use, so that the antibody will sink to the bottom of the tube to reach the indicated volume.

Q. Can I vortex a cell sample that has been labeled with an antibody?

   Yes, short-term shock can keep the cells in the best condition.

Q. Can cell samples labeled with tandem dyes such as APC-eFluor® 780, PE-Cy7 be fixed and stored?

   In most cases, samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL sample mix 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) , in the dark 4 ° C It can be stored for up to 3 days and has no effect on the test. Some effects may be related to the clone number.

Q. How can I save samples that cannot be detected immediately?

   First of all, to get the best results, it is recommended to check on the machine immediately.
To save the stained sample, you can save it in IC Fixation Buffer (cat. 00-8222) (100 μL sample mix 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) . At 4 ° C It can be stored for up to 3 days. For tandem dyes, such as APC-eFluor® 780, PE-Cy7, etc., eBioscience data shows no increase in their compensation with APC or PE . Some effects may be related to the clone number.

Q. Can I use my own fixed buffer to store cells?

   We do not recommend the use of other buffers, we can only ensure the preservation of IC Fixation Buffer (cat. 00-8222) or 1-step Fix/Lyse Solution (cat. 00-5333) .

Q. There are references to the incompatibility with DEC205+ cells . Which other markers can I choose?

   In fact, CD205 has been shown to bind to PE-Cy5.5- labeled antibodies and streptavidin markers (Part et al. 2012 J. Immunol Methods 384:184-90); however, it cannot be used for dendritic CD205+ cell. We recommend using PerCP-Cy5.5 or PerCP-eFluor 710 tags instead.

Q. Can I vortex an antibody or recombinant protein?

    We do not recommend oscillating proteins or antibody solutions, which can alter the characteristics of the product. It is usually possible that the liquid has already been dispersed in the pipe during transportation. It is recommended to centrifuge immediately before use, so that the antibody will sink to the bottom of the tube to reach the indicated volume.

Q. Can I vortex a cell sample that has been labeled with an antibody?

   Yes, short-term shock can keep the cells in the best condition.

Q. Can cell samples labeled with tandem dyes such as APC-eFluor® 780, PE-Cy7 be fixed and stored?

   In most cases, samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL sample mix 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) , in the dark 4 ° C It can be stored for up to 3 days and has no effect on the test. Some effects may be related to the clone number.

Q. How can I save samples that cannot be detected immediately?

   First of all, to get the best results, it is recommended to check on the machine immediately.
To save the stained sample, you can save it in IC Fixation Buffer (cat. 00-8222) (100 μL sample mix 100 μL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) . At 4 ° C It can be stored for up to 3 days. For tandem dyes, such as APC-eFluor® 780, PE-Cy7, etc., eBioscience data shows no increase in their compensation with APC or PE . Some effects may be related to the clone number.

Q. Can I use my own fixed buffer to store cells?

   We do not recommend the use of other buffers, we can only ensure the preservation of IC Fixation Buffer (cat. 00-8222) or 1-step Fix/Lyse Solution (cat. 00-5333) .

Q. There are references to the incompatibility with DEC205+ cells . Which other markers can I choose?

   In fact, CD205 has been shown to bind to PE-Cy5.5- labeled antibodies and streptavidin markers (Part et al. 2012 J. Immunol Methods 384:184-90); however, it cannot be used for dendritic CD205+ cell. We recommend using PerCP-Cy5.5 or PerCP-eFluor 710 tags instead.