For flow cytometry, blood cell lysis buffer (eBioscience, San Diego, CA, USA) was added to bone marrow cells, peripheral blood, and spleen cells of different groups of mice and incubated for 15 min at 4 °C. Then, cells were centrifuged and suspended in buffer containing 2% FBS in PBS and stained using the following fluorescently labeled antibodies purchased from BD Bioscience for 30 min: Ly6G-PE, CD11b-FITC, CD206-APC, and isotype controls (FITC, PE, and APC). After filtering through a 40-μm nylon cell strainer, samples were detected using an LSRFortessa SORP flow cytometer (BD, NJ, USA). Ly6G+CD11b+ cells were gated as neutrophils, and data analysis was implemented with FlowJo software (Ashland, OR, USA).
Blood cell lysis buffer
Manufactured by Thermo Fisher Scientific
Sourced in United States
Blood cell lysis buffer is a solution used to disrupt the cell membranes of blood cells, releasing their contents for further analysis. It is a core component in various blood and cell-based assays and procedures.
Automatically generated - may contain errors
2 protocols using blood cell lysis buffer
1
Neutrophil Profiling in Mouse Samples
2
Comprehensive Flow Cytometry Analysis of HL-60 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For ow cytometry analysis, blood cell lysis buffer (eBioscience, San Diego, CA, USA) was added to peripheral blood and bone marrow cells. Then, the cells were centrifuged and suspended in 2% fetal bovine serum (FBS) in PBS. Next, cells were stained with Ly-6G-PE, CD11b-PerCP/PECy5.5, CD16-FITC and CD206-APC (BD Biosciences, New Jersey, USA). After ltration through a 40 μm nylon cell strainer, samples were detected on a BD Accuri™ C6 for uorescence ow cytometry analysis.
Human HL-60 cell culture Human HL-60 cells (ATCC, Rockville, MD, USA) were cultured in RPMI-1640 medium containing 10% heatinactivated FBS and 1% penicillin-streptomycin at 37°C in a humidi ed atmosphere of 5% CO 2 /95% air.
Cells were divided into 3 groups: (1) Control group, (2) HL-60 + antagomiR-494 (50 nM) group, (3) HL-60 + antagomiR-494 (50 nM) + HDAC2 siRNA (50 nM) group. After cultivation for 24 hours, 200 µl supernatant was collected for further experiments, and the remaining cells were collected for the detection of HDAC2 mRNA, MMP7 mRNA, MMP10 mRNA, MMP13 mRNA, MMP16 mRNA, MMP21 mRNA, MMP25-AS1 mRNA, and MMP27 mRNA. Primers are listed in Table 1. Another set of transfected HL-60 cells was prepared for coculture with primary neurons.
Human HL-60 cell culture Human HL-60 cells (ATCC, Rockville, MD, USA) were cultured in RPMI-1640 medium containing 10% heatinactivated FBS and 1% penicillin-streptomycin at 37°C in a humidi ed atmosphere of 5% CO 2 /95% air.
Cells were divided into 3 groups: (1) Control group, (2) HL-60 + antagomiR-494 (50 nM) group, (3) HL-60 + antagomiR-494 (50 nM) + HDAC2 siRNA (50 nM) group. After cultivation for 24 hours, 200 µl supernatant was collected for further experiments, and the remaining cells were collected for the detection of HDAC2 mRNA, MMP7 mRNA, MMP10 mRNA, MMP13 mRNA, MMP16 mRNA, MMP21 mRNA, MMP25-AS1 mRNA, and MMP27 mRNA. Primers are listed in Table 1. Another set of transfected HL-60 cells was prepared for coculture with primary neurons.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!